Intelligent cancer diagnosis and treatment system and equipment based on combination of PI3K inhibitor GDC-0941 and diet restriction

Abstract

The invention discloses an intelligent cancer diagnosis and treatment system and equipment based on combination of a PI3K inhibitor GDC-0941 and diet restrictions, and finds and verifies that a treatment scheme of combination of the PI3K inhibitor GDC-0941 and the diet restrictions has a synergistic treatment effect on cancer treatment for the first time through experiments. The invention provides a brand-new intelligent cancer diagnosis and treatment system and equipment based on combination of a PI3K inhibitor GDC-0941 and diet restriction, and a computer readable storage medium. A brand-new thought and strategy are provided for research and development of a cancer treatment method.

Description

Technical Field

[0001] The present invention belongs to the field of intelligent diagnosis and treatment technology, and specifically relates to an intelligent cancer diagnosis and treatment system and equipment based on the combination of the PI3K inhibitor GDC-0941 and dietary restriction. Background Art

[0002] Phosphatidylinositol 3-kinase (PI3K) is a key enzyme responsible for regulating important processes such as cell proliferation, survival, and metabolism. It catalyzes the conversion of phosphatidylinositol (4,5)-bisphosphate (PIP2) to phosphatidylinositol (3,4,5)-triphosphate (PIP3), thereby activating downstream signaling pathways involving PDK1 and AKT, which are crucial for cell growth and survival. Aberrant activation of the PI3K signaling pathway is one of the most common oncogenic events in various human cancers. This aberrant signaling is often triggered by activating mutations in the PIK3CA gene (such as E545K or H1047R) or loss-of-function mutations in the tumor suppressor gene PTEN, making PI3K an attractive target for cancer treatment. Despite the solid theoretical basis for PI3K-targeted therapy in cancer treatment, the clinical efficacy of PI3K inhibitors (such as GDC-0941) has not met expectations. When used as monotherapy, these inhibitors generally exhibit limited antitumor activity, primarily through slowing proliferation by inducing cell cycle arrest rather than triggering cell death. This limited efficacy highlights a core challenge in cancer treatment: how to transform PI3K inhibitors from drugs that simply slow tumor growth into treatments that can effectively eradicate tumors. Addressing this issue is crucial to further enhance the potential of PI3K-targeted therapies in cancer treatment.

[0003] In recent years, non-apoptotic cell death mechanisms have attracted increasing attention, with this research direction aiming to address the challenges of drug resistance faced by traditional apoptosis-induced cancer therapies. Methuosis (derived from the Greek word metho, meaning "intoxication by alcohol") is a unique form of non-apoptotic cell death characterized by the accumulation of cytoplasmic vacuoles due to dysregulated macropinocytosis. Macropinocytosis is a process that uptakes extracellular fluid and nutrients through membrane ruffling and invagination. Although this mechanism normally helps cells survive nutrient-deprived conditions by taking up external nutrients, its dysregulation leads to widespread cytoplasmic vacuolation, which is unable to fuse with lysosomes, ultimately triggering lethal cell swelling, organelle displacement, and plasma membrane rupture. In the nutrient-restricted tumor microenvironment, due to insufficient vascularization and the high nutrient demands of rapidly proliferating cancer cells, tumor cells often upregulate macropinocytosis to meet their metabolic needs. This dependence on macropinocytosis creates a therapeutic vulnerability that can be exploited as a strategy to selectively induce cancer cell death. However, few studies have investigated nutrient restriction as a strategy to amplify the cytotoxic effects of methuosis-inducing agents by exploiting the dependence of cancer cells on macropinocytosis. Summary of the Invention

[0004] In view of this, in order to overcome the deficiencies in the above-mentioned prior art, the present invention provides a cancer intelligent diagnosis and treatment system and device based on the combination of the PI3K inhibitor GDC-0941 and dietary restriction.

[0005] This study reports for the first time that combining PI3K inhibition with nutrient restriction can induce methuosis in cancer cells with abnormally activated PI3K. By using a PI3K inhibitor under conditions that mimic nutrient deprivation, we induced dysregulated macropinocytosis, triggering cytotoxic methuosis-like cell death in both in vitro and in vivo models. This selective cell-killing effect redefines the mechanism of action of GDC-0941, transforming it from a drug that primarily inhibits cell proliferation to a therapeutic with potent cytotoxicity. This study provides a novel therapeutic strategy for cancer treatment, combining a PI3K inhibitor with dietary restriction, expanding the application prospects of PI3K-targeted therapies.

[0006] The present invention adopts the following technical solutions to achieve the above-mentioned invention objectives:

[0007] The first aspect of the present invention provides a cancer intelligent diagnosis and treatment system based on the combination of PI3K inhibitor GDC-0941 and dietary restriction.

[0008] Furthermore, the system comprises:

[0009] A diagnostic element information collector is used to collect cancer diagnostic element information related to the patient and send the collected data results to the disease diagnosis matcher;

[0010] A disease diagnosis matcher, configured to match the diagnosis element information from the diagnosis element information collector with the cancer type, and send the matched data results to a treatment plan matcher;

[0011] A treatment plan matcher, configured to determine the matching result of the disease diagnosis matcher and call a treatment plan according to the matching result, and then send the treatment plan data result to the disease diagnosis and treatment information outputter;

[0012] A disease diagnosis and treatment information output device, configured to output the data results received from the disease diagnosis matcher and the treatment plan matcher to a receiving unit;

[0013] In the treatment plan matcher, if the disease diagnosis matcher matches the patient to be diagnosed as a cancer patient, the following treatment plan is called: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restrictions to the patient to be diagnosed;

[0014] In the treatment plan matcher, if the disease diagnosis matcher matches the patient to be diagnosed as a non-cancer patient, the following treatment plan is called without matching: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient.

[0015] Furthermore, the cancer diagnosis elements include medical history, symptom manifestations, examination data, imaging examination data, laboratory examination data and / or pathological examination data;

[0016] Optionally, the cancer is a cancer in which the PI3K signaling pathway is altered.

[0017] Optionally, the cancer is ovarian cancer, cervical cancer, breast cancer or pancreatic cancer.

[0018] Furthermore, in the disease diagnosis matching device, if the examination data shows that a tumor or mass exists in the relevant tissue, the imaging examination data shows that a cystic or solid mass exists in the relevant tissue, the laboratory examination data shows that the CA125 value in the patient's serum is higher than the normal range, and / or the pathological examination data shows that the biopsy tissue is cancerous tissue, the patient is matched as a cancer patient;

[0019] In the disease diagnosis matching device, if the examination data shows that there is no abnormality in the relevant tissue, the imaging examination data shows that there is no abnormality, the laboratory examination data shows that the CA125 value in the serum of the patient is within the normal range and / or the pathological examination data shows that the biopsy tissue is normal tissue, the patient is matched as a non-cancer patient.

[0020] Furthermore, the combination of the PI3K inhibitor GDC-0941 and dietary restriction in the treatment regimen has a synergistic therapeutic effect on the treatment of cancer, especially for the treatment of ovarian cancer, cervical cancer, breast cancer or pancreatic cancer.

[0021] Furthermore, the receiving unit is a display screen, a computer client, a mobile phone client or a tablet.

[0022] In some embodiments, the cancer is ovarian cancer, cervical cancer, breast cancer, or pancreatic cancer, and can be diagnosed using common examination methods and diagnostic criteria for ovarian cancer, cervical cancer, breast cancer, or pancreatic cancer known to those skilled in the art.

[0023] In one embodiment, if the examination data shows that there are tumors or masses in the ovarian, cervical, breast or pancreatic tissues, the imaging examination data shows that there are cystic or solid masses in the ovarian, cervical, breast or pancreatic tissues, the laboratory examination data shows that the CA125 value in the patient's serum is higher than the normal range and / or the pathological examination data shows that the biopsy tissue is ovarian cancer, cervical cancer, breast cancer or pancreatic cancer tissue, then the patient is matched as a cancer patient.

[0024] In one embodiment, if the examination data shows that there is no abnormality in the ovarian, cervical, breast or pancreatic tissue, the imaging examination data shows that there is no abnormality, the laboratory examination data shows that the CA125 value in the patient's serum is within the normal range and / or the pathological examination data shows that the biopsy tissue is normal ovarian, cervical, breast or pancreatic tissue, the patient is matched as a non-cancer patient.

[0025] The second aspect of the present invention provides an intelligent cancer diagnosis and treatment device based on the combination of PI3K inhibitor GDC-0941 and dietary restriction.

[0026] Furthermore, the device includes a memory and a processor; the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, it is used to perform the following method:

[0027] S101: Obtaining cancer diagnosis element information related to the patient;

[0028] S102: Determining whether the patient is a cancer patient based on the cancer diagnosis element data related to the patient;

[0029] S103: When the patient is a cancer patient, the following treatment regimen is selected: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient.

[0030] Furthermore, the cancer diagnosis elements include medical history, symptom manifestations, examination data, imaging examination data, laboratory examination data and / or pathological examination data;

[0031] Optionally, if the examination data shows that a tumor or mass exists in the relevant tissue, the imaging examination data shows that a cystic or solid mass exists in the relevant tissue, the laboratory examination data shows that the CA125 value in the patient's serum is higher than the normal range, and / or the pathological examination data shows that the biopsy tissue is cancerous tissue, then the patient is determined to be a cancer patient;

[0032] Optionally, the cancer is a cancer in which the PI3K signaling pathway is altered.

[0033] Optionally, the cancer is ovarian cancer, cervical cancer, breast cancer or pancreatic cancer.

[0034] A third aspect of the present invention provides a computer-readable storage medium having a computer program stored thereon.

[0035] Furthermore, when the computer program is executed by a processor, the following method is implemented:

[0036] S101: Obtaining cancer diagnosis element information related to the patient;

[0037] S102: Determining whether the patient is a cancer patient based on the cancer diagnosis element data related to the patient;

[0038] S103: When the patient is a cancer patient, selecting the following treatment plan: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient;

[0039] Optionally, the cancer is a cancer in which the PI3K signaling pathway is altered.

[0040] Optionally, the cancer is ovarian cancer, cervical cancer, breast cancer or pancreatic cancer.

[0041] A fourth aspect of the present invention provides a method for inducing non-apoptotic cell death (methuosis) in cancer cells.

[0042] Furthermore, the method includes treating cancer cells with the PI3K inhibitor GDC-0941 in combination with dietary restriction;

[0043] Optionally, the cancer cells are ovarian cancer cells, cervical cancer cells, breast cancer cells or pancreatic cancer cells.

[0044] A fifth aspect of the present invention provides a system for inducing non-apoptotic cell death (methuosis) in cancer cells.

[0045] Furthermore, the system comprises:

[0046] an acquisition unit, used for acquiring cancer cells to be processed;

[0047] a treatment unit, treating the cancer cells with the PI3K inhibitor GDC-0941 in combination with dietary restriction to induce methuosis;

[0048] Optionally, the cancer cells are ovarian cancer cells, cervical cancer cells, breast cancer cells or pancreatic cancer cells.

[0049] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0050] The present invention discovered for the first time and experimentally confirmed that the treatment regimen of the PI3K inhibitor GDC-0941 combined with dietary restriction has a synergistic therapeutic effect in the treatment of cancer (especially ovarian cancer, cervical cancer, breast cancer or pancreatic cancer). Based on this, the present invention provides a new cancer intelligent diagnosis and treatment system, equipment and computer-readable storage medium based on the combination of the PI3K inhibitor GDC-0941 and dietary restriction. The present invention provides a new idea and strategy for the research and development of cancer treatment methods, and has important scientific significance and clinical application value in the technical field of cancer treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 : Schematic diagram of an intelligent cancer diagnosis and treatment system based on the combination of the PI3K inhibitor GDC-0941 and dietary restriction, provided by an embodiment of the present invention;

[0052] Figure 2 : Schematic diagram of an intelligent cancer diagnosis and treatment device based on the combination of PI3K inhibitor GDC-0941 and dietary restriction, provided by an embodiment of the present invention;

[0053] Figure 3 : Experimental results showing that under serum starvation conditions, PI3K inhibition induces giant cytoplasmic vacuoles. GDC-0941-induced vacuoles exhibit characteristics of macropinosomes.

[0054] Figure 4 : The results corresponding to the enhanced cell death induced by cytoplasmic vacuolization induced by PI3K inhibitor GDC-0941;

[0055] Figure 5 : The corresponding result diagram shows that the conversion of PIP2 is crucial for GDC-0941-induced methuosis;

[0056] Figure 6 :PI3K inhibitor GDC-0941 selectively induces methuosis in PI3K-dependent cancer cells;

[0057] Figure 7: PI3K inhibitor GDC-0941 induces methuosis in multiple cancer cell lines through PI3K-independent macropinocytosis;

[0058] Figure 8 : The corresponding results of the PI3K inhibitor GDC-0941 and dietary restriction synergize to induce methuosis and inhibit tumor growth in vivo. DETAILED DESCRIPTION

[0059] The present invention will be further described below in conjunction with specific examples. The following specific examples are only used to explain the present invention and are not to be construed as limiting the present invention. Those of ordinary skill in the art will appreciate that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and purpose of the present invention, and that the scope of the present invention is defined by the claims and their equivalents. The reagents and raw materials used in the present invention are readily available to those of ordinary skill in the art and are commercially available unless otherwise specified. The experimental methods for which specific conditions are not specified in the present invention are typically tested under conventional conditions or under conditions recommended by the manufacturer. In particular, the following examples are only used to illustrate the present invention and are not to be construed as limiting the scope of the present invention in any way. It should be noted that the experimental conditions and results described in the following examples are only used to illustrate the present invention and should not and will not limit the present invention described in detail in the claims.

[0060] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. In some of the processes described in the specification and claims of the present invention and the above-mentioned drawings, multiple operations that appear in a specific order are included, but it should be clearly understood that these operations may not be executed in the order in which they appear in this document or may be executed in parallel. The serial numbers of the operations, such as S101, S102, S103, etc., are only used to distinguish between different operations, and the serial numbers themselves do not represent any execution order. In addition, these processes may include more or fewer operations, and these operations may be executed sequentially or in parallel.

[0061] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making any creative efforts shall fall within the scope of protection of the present invention.

[0062] Figure 1Schematic diagram of an intelligent cancer diagnosis and treatment system based on the combination of the PI3K inhibitor GDC-0941 and dietary restriction, provided in an embodiment of the present invention. The system includes:

[0063] A diagnostic element information collector is used to collect cancer diagnostic element information related to the patient and send the collected data results to the disease diagnosis matcher;

[0064] A disease diagnosis matcher, used to match the diagnosis element information of the diagnosis element information collector with the disease type, and send the matched data results to the treatment plan matcher;

[0065] A treatment plan matcher, configured to determine the matching result of the disease diagnosis matcher and call a treatment plan according to the matching result, and then send the treatment plan data result to the disease diagnosis and treatment information outputter;

[0066] A disease diagnosis and treatment information output device, configured to output the data results received from the disease diagnosis matcher and the treatment plan matcher to a receiving unit;

[0067] In the treatment plan matcher, if the disease diagnosis matcher matches the patient to be diagnosed as a cancer patient, the following treatment plan is called: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restrictions to the patient to be diagnosed;

[0068] In the treatment plan matcher, if the disease diagnosis matcher matches the patient to be diagnosed as a non-cancer patient, the following treatment plan is called without matching: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient.

[0069] In some embodiments, the cancer diagnosis elements include medical history, symptom manifestations, examination data, imaging examination data, laboratory examination data and / or pathological examination data.

[0070] In some embodiments, in the disease diagnosis matching device, if the examination data shows that a tumor or mass exists in the relevant tissue, the imaging examination data shows that a cystic or solid mass exists in the relevant tissue, the laboratory examination data shows that the CA125 value in the patient's serum is higher than the normal range, and / or the pathological examination data shows that the biopsy tissue is cancerous tissue, the patient is matched as a cancer patient;

[0071] In the disease diagnosis matching device, if the examination data shows that there is no abnormality in the relevant tissue, the imaging examination data shows that there is no abnormality, the laboratory examination data shows that the CA125 value in the serum of the patient is within the normal range and / or the pathological examination data shows that the biopsy tissue is normal tissue, the patient is matched as a non-cancer patient.

[0072] In some embodiments, the cancer diagnostic factors are not limited to the above-mentioned medical history, symptom manifestations, examination data, imaging examination data, laboratory examination data and / or pathological examination data. Any factors that can be used to diagnose cancer known to those skilled in the art are included in the cancer diagnostic factors described in the present invention. In addition, those skilled in the art can diagnose whether the subject has cancer based on the above-mentioned cancer diagnostic factors and diagnostic criteria known in the art.

[0073] It should be noted that the innovation of the present invention lies not in the method for diagnosing cancer, but in the treatment plan adopted for cancer patients: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restrictions to the patient (i.e., cancer patient).

[0074] In a specific embodiment of the present invention, the cancer patient is a cancer patient with an altered PI3K signaling pathway, specifically, the cancer patient is an ovarian cancer patient, a cervical cancer patient, a breast cancer patient, or a pancreatic cancer patient.

[0075] In some embodiments, the effective amount refers to an amount with a therapeutic effect or the amount required to produce a therapeutic effect in a subject. For example, a pharmaceutically effective amount refers to the amount of the drug required to produce the desired therapeutic effect, and the therapeutic effect can be reflected by the results of clinical trials, model animal studies and / or in vitro studies. The pharmaceutically effective amount depends on several factors, including but not limited to: characteristic factors of the subject (such as height, weight, sex, age and medication history), the type of disease, and the severity of the disease. Clinicians can determine the amount of PI3K inhibitor GDC-0941 required to produce a therapeutic effect in a subject based on the above factors of the specific subject.

[0076] In some embodiments, the PI3K inhibitor GDC-0941 is a novel selective class I phosphatidylinositol 3-kinase (PI3K) inhibitor, the corresponding CAS number of which is 957054-30-7, and the corresponding molecular formula of which is C 23 H 27 N7O3S2, with a corresponding molecular weight of 513.64. Activation of the PI3K / Akt signaling pathway is frequently associated with tumorigenesis. Dysregulation of this pathway frequently occurs in various cancers and may lead to resistance to many anticancer agents. GDC-0941 competitively binds to the ATP-binding pocket of PI3K, preventing the formation of phosphatidylinositol (3,4,5)-triphosphate (PIP3), a second messenger that transmits downstream PI3K signals.

[0077] Although GDC-0941 is generally well tolerated with manageable side effects, early clinical trials have shown that its efficacy as a monotherapy is limited, mainly manifested as inhibition of cell proliferation rather than induction of cytotoxicity. These findings highlight the need to develop more effective treatment strategies. In this study, the inventors demonstrated that GDC-0941 can induce a form of non-apoptotic cell death, Methuosis, whose effects are significantly enhanced under nutrient-restricted conditions. The notable feature of Methuosis is cytoplasmic vacuolization driven by dysregulated macropinocytosis. These vacuoles have a single membrane structure and co-localize with early and late endosomal markers (EEA1 and Rab7, respectively). By using BafA1 (vacuolar H + -ATPase inhibitor) or EIPA (macropinocytosis inhibitor) blocked vacuolar formation, further verifying the key role of macropinocytosis in this process. Importantly, these vacuoles cannot fuse with lysosomes, thereby disrupting the normal macropinocytosis pathway. In addition, vacuolation can be partially reversed after GDC-0941 withdrawal, indicating that continued inhibition of PI3K is crucial for maintaining its cytotoxic effect. The inventors also found that under nutrient-restricted conditions, GDC-0941 significantly promoted the occurrence of Methuosis. Serum starvation enhanced the extent of vacuolation and cell death, especially in PI3K-dependent cancer cells (such as OVSAHO and SNU119). In contrast, PI3K-independent cells, such as OVCAR4, showed resistance. However, introducing the PIK3CA E545K mutation into OVCAR4 cells allowed them to obtain sustained PI3K activation, making these cells more susceptible to GDC-0941-induced Methuosis under serum starvation conditions. These findings highlight the selective role of methuosis in PI3K-dependent cancers under nutrient-restricted conditions, providing new avenues for PI3K-targeted therapy and highlighting the potential for enhancing therapeutic efficacy by combining nutrient restriction strategies.

[0078] In some embodiments, the dietary restriction is caloric restriction.

[0079] In a specific embodiment of the present invention, dietary restriction (DR) refers to controlling the total daily caloric intake of mice to only 60% of the caloric intake of the adlibitum (AL) group. To prevent malnutrition due to caloric reduction, the feed of the DR group is fortified with vitamins and minerals, while maintaining a similar composition and form to that of the adlibitum group.

[0080] In some embodiments, the combination of the PI3K inhibitor GDC-0941 and dietary restriction in the treatment regimen has a synergistic therapeutic effect on the treatment of cancer (especially ovarian cancer, cervical cancer, breast cancer or pancreatic cancer).

[0081] In some embodiments, the receiving unit is a display screen, a computer client, a mobile phone client or a tablet.

[0082] In a specific embodiment, the present invention demonstrates that under serum starvation conditions, PI3K inhibition induces giant cytoplasmic vacuoles, and demonstrates that GDC-0941-induced vacuoles exhibit characteristics of macropinocytic bodies. Figure 3 Specifically, as follows:

[0083] (1) Experimental methods

[0084] 1) Drug Treatment and Morphological Observation: OVSAHO and SNU119 cells (ovarian cancer cells) were treated with the PI3K inhibitor GDC-0941 or the dual PI3K / mTOR inhibitor BEZ235 for 24 hours in standard culture medium supplemented with 10% fetal bovine serum or serum-free culture medium, respectively. After treatment, cell morphological changes were observed under a microscope and photographed.

[0085] 2) Western blot analysis: Cells were lysed with ice-cold RIPA buffer containing protease and phosphatase inhibitors (Roche), and total protein was extracted. Proteins were separated by SDS-PAGE electrophoresis, transferred to a membrane, and incubated with primary antibodies overnight at 4°C. Images were analyzed using fluorescent secondary antibodies and the Odyssey infrared imaging system (Li-COR).

[0086] 3) Transmission Electron Microscopy Analysis: OVSAHO cells were serum-starved and treated with GDC-0941 for 24 hours before being fixed with 2.5% glutaraldehyde (pH 7.4). After fixation, cells were washed with 0.1 M phosphate buffer and fixed with 1% molybdenum tetroxide for 2 hours. Cells were then dehydrated through a graded ethanol series, embedded in resin, and sectioned. Images were captured using a JEM-1400 transmission electron microscope.

[0087] 4) Live Cell Imaging: To observe vacuolation and endocytosis, cells were treated with or without GDC-0941 for 24 hours. Lucifer Yellow (LY, 1 mg / mL, Sigma-Aldrich, Catalog No. L0144) was then added and incubated at 37°C for 2 hours to label intracellular vacuoles. Intracellular acidic structures were stained with 50 nM Lyso-Tracker Red (ThermoFisher Scientific, Catalog No. L12492) for 30 minutes. Images were acquired using a Leica TCS SP8 confocal microscope, and cell morphology was observed using a Leica DMi1 inverted microscope. Four fields of view were randomly selected for image analysis for each treatment condition.

[0088] 5) Immunofluorescence microscopy: After fixation, permeabilization, and blocking, cells were incubated with primary antibodies overnight at 4°C. Primary antibodies included EEA1 (Santa Cruz, sc-137130) and Rab7 (CST, #95746). Fluorescently labeled secondary antibodies (Thermo Fisher Scientific) and DAPI staining solution (Sigma-Aldrich) were then added for staining. Fluorescence images were acquired using a Leica DM6B Thunder fluorescence microscope.

[0089] (2) Experimental results

[0090] The pan-class I PI3K inhibitor GDC-0941 has been shown to exhibit antiproliferative effects in various cancer types. In this study, we found that treatment with GDC-0941 under standard culture conditions induced the formation of numerous cytoplasmic vacuoles in the ovarian cancer cell lines OVSAHO and SNU119. To prevent interference of serum growth factors with PI3K / AKT signaling, we treated cells with GDC-0941 in the absence of serum. Results showed that under these conditions, GDC-0941 not only induced vacuoles but also promoted the formation of larger cytoplasmic vacuoles, suggesting that nutrient restriction enhances the cellular response to PI3K inhibition. To elucidate the origin of GDC-0941-induced cytoplasmic vacuoles, we used transmission electron microscopy to investigate their ultrastructure. Following 24 hours of serum-starved treatment with GDC-0941, numerous single-membrane vacuoles, devoid of cytoplasmic components or organelles, appeared in OVSAHO cells. These vacuoles were distinct from autophagosomes, which typically contain cytoplasmic components and have a double membrane. Another key observation was the presence of irregular plasma membrane protrusions and invaginations, features consistent with macropinocytosis.

[0091] To confirm that these cytoplasmic vacuoles originated from macropinosomes, we treated OVSAHO and SNU119 cells with GDC-0941 for 24 hours under serum starvation conditions and then added Lucifer yellow (a liquid phase tracer of macropinosomes) for incubation. Subsequently, confocal microscopy was used to observe. The results showed that most vacuoles were Lucifer yellow positive, supporting their characteristics as macropinosomes. In addition, these vacuoles can be marked by the early endosome marker EEA1 and the late endosome marker Rab7. Using vacuolar-type H + GDC-0941-induced vacuolization was effectively inhibited by treatment with the ATPase inhibitor Bafilomycin A1 (BafA1), which is essential for the macropinocytosis process. 5-(N-ethyl-N-isopropyl)amiloride (EIPA) is a Na + / H + Exchange inhibitors, which have also been shown to effectively block macropinocytic body formation, also reversed GDC-0941-induced vacuolization. These results further confirmed that GDC-0941-induced cytoplasmic vacuoles originated from macropinocytic bodies. In addition, Lyso-Tracker Red labeling showed that there was no colocalization between GDC-0941-induced vacuoles and lysosomes, indicating that these vacuoles did not fuse with lysosomes. These findings strongly suggest that the PI3K inhibitor GDC-0941 induces vacuolization in cancer cells under nutrient-deprived conditions by inducing dysregulated macropinocytosis.

[0092] In a specific embodiment, the present invention demonstrates that GDC-0941-induced cytoplasmic vacuolization enhances cell death, as shown in the experimental results. Figure 4 Specifically, as follows:

[0093] (1) Experimental methods

[0094] 1) Autophagy Flux Assay: A cancer cell line stably expressing mRFP1-EGFP-LC3B was constructed using the lentiviral vector pCDH-CMV-mRFP1-EGFP-LC3B-EF1-Puro (Miaoling Biotechnology, Cat. No. P4838). Stable cell lines were obtained after infection by puromycin selection. After treatment with the corresponding drugs, cells were fixed with 4% paraformaldehyde and imaged under a Leica DM6B Thunder fluorescence microscope. At least 30 cells were analyzed per group, and the number of yellow (mRFP+EGFP) and red (mRFP-single-positive) puncta in each cell was counted to assess autophagy flux levels.

[0095] 2) Western blot analysis: Cells were lysed with ice-cold RIPA buffer containing protease and phosphatase inhibitors (Roche), and total protein was extracted. Proteins were separated by SDS-PAGE electrophoresis, transferred to a membrane, and incubated with primary antibodies overnight at 4°C. Images were analyzed using fluorescent secondary antibodies and the Odyssey infrared imaging system (Li-COR).

[0096] 3) Flow cytometry: To detect cell death, propidium iodide (PI, Sigma) staining was performed. The steps were as follows: cells were harvested, washed with PBS, and then PI staining solution was added. The staining was performed according to the reagent instructions to assess cell membrane integrity. The stained cells were analyzed using a BD FACS Canto II flow cytometer (BD Biosciences).

[0097] (2) Experimental results

[0098] We examined whether GDC-0941-induced vacuolization affects cell viability. Under serum-starved conditions, GDC-0941 treatment initiated the formation of cytoplasmic vacuoles, which gradually increased in size over time, ultimately leading to cell membrane rupture and cell death, effectively inhibiting cell growth. Notably, pretreatment with the apoptosis inhibitor Z-VAD-FMK did not reduce vacuolization or prevent cell death, confirming that GDC-0941-induced cell death is non-apoptotic. These results suggest that GDC-0941 impairs cancer cell growth and survival through a vacuole-mediated cell death mechanism.

[0099] Inhibition of the PI3K / AKT / mTOR signaling pathway is known to trigger autophagy, a cellular process that helps cancer cells survive under nutrient and energy stress. Using an mRFP-GFP-LC3 tandem fluorescent plasmid to monitor autophagic flux, we observed that GDC-0941 treatment significantly increased both yellow (representing colocalization of mRFP and GFP) and red puncta (representing mRFP alone, due to quenching of GFP in acidic lysosomes). These findings suggest enhanced autophagic flux. Furthermore, GDC-0941 treatment increased LC3-II / LC3-I levels and decreased p62 / SQSTM1 (an autophagy adaptor) levels in cells. These results collectively indicate that GDC-0941 induces autophagy in cancer cells. Next, we investigated the effects of autophagy on GDC-0941-induced vacuolation and cell growth and survival using the autophagy inhibitor MRT68921, a drug that inhibits autophagy initiation. We found that MRT68921 had minimal effect on GDC-0941-induced vacuolation.

[0100] Notably, GDC-0941-induced vacuolation was reversible after drug removal, as evidenced by the disappearance of vacuoles and restoration of cell viability. Overall, these results highlight the importance of continuous GDC-0941 treatment as a therapeutic approach to induce methuotic cell death.

[0101] In a specific embodiment, the present invention demonstrates that the conversion of PIP2 is crucial for GDC-0941-induced methuosis. The experimental results are as follows: Figure 5 Specifically, as follows:

[0102] (1) Experimental methods

[0103] 1) RNA sequencing and proteomic analysis

[0104] RNA sequencing analysis: Ovarian cancer cell lines OVSAHO and SNU119 were treated with 5 μM GDC-0941 for 24 hours or without treatment under serum-free conditions. RNA was extracted using TRIzol reagent and sent to Novogene for sequencing. Ultra TM RNA library was constructed using RNA Library Prep Kit, and the molecular functional pathways related to GDC-0941 treatment were analyzed by Gene Set Enrichment Analysis (GSEA), with significance thresholds set at p < 0.01 and FDR < 0.05.

[0105] Proteomic Analysis: Cells treated with 5 μM GDC-0941 for 24 hours or left untreated were lysed with lysis buffer, and proteins were extracted and digested with trypsin. Analysis was performed by LC-MS / MS, with peptides separated using a C18 column. Data were acquired in DIA mode and aligned to the UniProt-Human database using Spectronaut 19 software.

[0106] Data Analysis: Peptides identified by mass spectrometry or transcripts with a log2 fold change > 0.5 and p < 0.05 were considered differentially expressed genes or proteins. GO cellular component enrichment analysis was performed using ClusterProfiler.

[0107] 2) Quantitative reverse transcription PCR (qRT-PCR)

[0108] Total RNA was extracted using TRIzol reagent (Invitrogen) according to the manufacturer's instructions, and reverse transcribed into complementary DNA (cDNA) using the PrimeScript RT kit (Takara Bio, Japan). qRT-PCR was performed using the StepOnePlus Real-Time PCR System (Applied Biosystems) using SYBR Green PCR MasterMix (Applied Biosystems). Primers were synthesized by Sangon Biotech (China).

[0109] 3) siRNA transfection

[0110] siRNA oligonucleotides were transfected into cells using Lipofectamine 3000 (Invitrogen) according to the manufacturer's instructions. siRNA reagents were provided by GenePharma.

[0111] 4) Immunofluorescence microscopy: After fixation, permeabilization, and blocking, cells were incubated with primary antibodies overnight at 4°C. The primary antibody used was PIP2 (ab11039, Abcam). Fluorescently labeled secondary antibodies (Thermo Fisher Scientific) and DAPI staining solution (Sigma-Aldrich) were then added for staining. Fluorescence images were acquired using a Leica DM6B Thunder fluorescence microscope.

[0112] 5) PIP2 Distribution Assay: To detect intracellular PIP2 distribution, cells were transfected with a GFP-tagged PLCδPH domain expression plasmid (pLV3-CMV-EGFP-PLCδ-PH-Puro, Miaoling Biotechnology, Cat. No. P50541) using Lipofectamine 3000 (Invitrogen) for 48 hours. Subsequently, cells were treated or untreated with GDC-0941 in serum-free conditions for 24 hours and fixed with 4% paraformaldehyde. GFP signals were observed using a Leica DM6B Thunder fluorescence microscope to analyze the cellular distribution of PIP2.

[0113] 6) ROS Detection: To measure intracellular reactive oxygen species (ROS) levels, cells were incubated with 5 μM H2DCFDA probe (Invitrogen, Cat. No. D399) at 37°C for 1 hour. After incubation, cells were washed with PBS, and fluorescence intensity was measured using a BD FACS Canto II flow cytometer to assess ROS levels.

[0114] (2) Experimental results

[0115] To further reveal the molecular basis of GDC-0941-induced methuosis, we performed integrated proteomic and transcriptomic analyses of OVSAHO and SNU119 cells following GDC-0941 treatment. Venn diagram analysis revealed that a total of 42 genes were significantly upregulated at both the mRNA and protein levels in both ovarian cancer cell lines. Pathway enrichment analysis of these genes revealed that they were closely associated with vacuolar-related structures (e.g., vacuolar lumen and cytoplasmic vesicle lumen), consistent with the methuosis phenotype. GSEA further revealed that these treated cells were significantly enriched for ion channel-related functions, including ligand-gated single-atom ion channel activity and gated channel activity, suggesting that ion channel dysfunction may play a role in GDC-0941-induced methuosis.

[0116] Given that phosphatidylinositol (4,5)-bisphosphate (PIP2) is a substrate of PI3K and regulates ion channels and membrane dynamics, we proposed that PIP2 accumulation upon PI3K inhibition might drive vacuole formation. Using the GFP-tagged phospholipase C delta (PLCδ) PH domain as a PIP2 probe, we observed that GDC-0941 treatment resulted in the relocalization of PIP2 from the plasma membrane to the vacuole membrane, a finding confirmed by PIP2 immunostaining. To further confirm the critical role of PIP2, we used the PLC agonist m-3M3FBS to promote PIP2 hydrolysis, resulting in a significant reduction in vacuole formation. Because methuosis involves excessive fluid uptake through the macropinocytosis pathway, a process that relies on water and ion channels to maintain cell and vesicle volume, we examined the role of the aquaporin AQP1 and chloride channels. Knockdown of AQP1 or inhibition of Cl- channels with NPPB significantly inhibited vacuole formation. In summary, the above results indicate that GDC-0941 promotes the accumulation and redistribution of PIP2, disrupting ion transport and cellular homeostasis, thereby inducing abnormal vacuolization and ultimately leading to methuosis.

[0117] In a specific embodiment, the present invention demonstrates that GDC-0941 selectively induces methuosis of PI3K-dependent cancer cells. The experimental results are as follows: Figure 6 Specifically, as follows:

[0118] (1) Experimental methods

[0119] 1) Western blot analysis: Cells were lysed with ice-cold RIPA buffer containing protease and phosphatase inhibitors (Roche), and total protein was extracted. Proteins were separated by SDS-PAGE electrophoresis, transferred to a membrane, and incubated with primary antibodies overnight at 4°C. Images were analyzed using fluorescent secondary antibodies and the Odyssey infrared imaging system (Li-COR).

[0120] 2) Live Cell Imaging: To observe vacuolation and endocytosis, cells were treated with or without GDC-0941 for 24 hours. Lucifer Yellow (LY, 1 mg / mL, Sigma-Aldrich, Catalog No. L0144) was then added and incubated at 37°C for 2 hours to label intracellular vacuoles. Intracellular acidic structures were stained with 50 nM Lyso-Tracker Red (ThermoFisher Scientific, Catalog No. L12492) for 30 minutes. Images were acquired using a Leica TCS SP8 confocal microscope, and cell morphology was observed using a Leica DMi1 inverted microscope. Four fields of view were randomly selected for image analysis for each treatment condition.

[0121] 3) Flow cytometry: To detect cell death, propidium iodide (PI, Sigma) staining was performed. The steps were as follows: cells were harvested, washed with PBS, and then PI staining solution was added. The staining was performed according to the reagent instructions to assess cell membrane integrity. The stained cells were analyzed using a BD FACS Canto II flow cytometer (BD Biosciences).

[0122] 4) Immunofluorescence microscopy: After fixation, permeabilization, and blocking, cells were incubated with primary antibodies overnight at 4°C. The primary antibody used was PIP2 (ab11039, Abcam). Fluorescently labeled secondary antibodies (Thermo Fisher Scientific) and DAPI staining solution (Sigma-Aldrich) were then added for staining. Fluorescence images were acquired using a Leica DM6B Thunder fluorescence microscope.

[0123] 5) PIP2 Distribution Assay: To detect intracellular PIP2 distribution, cells were transfected with a GFP-tagged PLCδPH domain expression plasmid (pLV3-CMV-EGFP-PLCδ-PH-Puro, Miaoling Biotechnology, Cat. No. P50541) using Lipofectamine 3000 (Invitrogen) for 48 hours. Subsequently, cells were treated or untreated with GDC-0941 in serum-free conditions for 24 hours and fixed with 4% paraformaldehyde. GFP signals were observed using a Leica DM6B Thunder fluorescence microscope to analyze the cellular distribution of PIP2.

[0124] (2) Experimental results

[0125] To determine whether sustained PI3K activation affects GDC-0941-induced methuosis in the above cancer cell lines, we constructed cells stably expressing oncogenic mutant PIK3CA. E545K The OVCAR4 ovarian cancer cell line was tested. This mutation is one of the common PI3K signaling drivers in cancer. Under serum starvation conditions, OVCAR4 cells expressing the empty vector showed significantly reduced basal levels of AKT phosphorylation compared to cells cultured in normal medium, while cells expressing PIK3CA E545K OVCAR4 cells expressing PIK3CA maintain a state of sustained AKT activation. This strategy allows us to specifically investigate how sustained PI3K signaling affects the cellular response to PI3K inhibition and the mechanism of induction of methuotic cell death under nutrient restriction conditions. E545K OVCAR4 cells expressing GDC-0941 showed extensive cytoplasmic vacuolation, while OVCAR4 cells expressing the empty vector produced almost no vacuoles. We confirmed that the vacuoles induced by GDC-0941 originated from abnormally activated macropinocytic bodies by detecting Lucifer yellow, and then found that such vacuoles eventually led to cell death by PI staining. + -ATPase inhibitor Bafilomycin A1 can completely block vacuole formation, indicating that vacuole acidification is crucial in the process of methuosis. Further analysis showed that GDC-0941 can inhibit PIK3CA E545K In cells expressing PLC, PIP2 was specifically redistributed to the vacuole membrane, a phenomenon observed by imaging with a GFP-PLCδ-PH probe and PIP2 immunostaining. Activation of PLC and hydrolysis of PIP2 by m-3M3FBS almost completely abolished vacuole formation, further confirming the critical role of PIP2 accumulation and redistribution in this process. In summary, ovarian cancer cells with aberrant PI3K activation were more sensitive to GDC-0941-induced methuosis under nutrient-deprived conditions, which may be related to their enhanced dependence on PI3K-mediated PIP2 metabolism. This finding suggests that combining PI3K inhibitors with nutrient restriction may have potential therapeutic value for PI3K-driven cancers.

[0126] In a specific embodiment, the present invention demonstrates that GDC-0941 induces methuosis in multiple cancer cell lines through PI3K-independent macropinocytosis. Figure 7 Specifically, as follows:

[0127] (1) Experimental methods

[0128] 1) Dextran uptake experiment: Cells were seeded on glass slides in 24-well plates. 24 hours after seeding, the cells were treated with or without GDC-0941 in serum-free medium for 24 hours. In order to label macropinocytic vesicles, a high molecular weight TMR-dextran (D1818, Thermo Fisher Scientific) uptake experiment was performed. TMR-dextran was added to serum-free medium at a final concentration of 1 mg / mL and incubated at 37°C for 30 minutes. After incubation, the cells were washed five times with cold PBS and immediately fixed with 3.7% formalin. The fixed cells were stained with DAPI in PBS for 10 minutes and then washed three times with PBS. Fluorescence images were taken using a DM6B Thunder fluorescence microscope (Leica Microsystems).

[0129] 2) PIP2 Distribution Assay: To detect intracellular PIP2 distribution, cells were transfected with a GFP-tagged PLCδPH domain expression plasmid (pLV3-CMV-EGFP-PLCδ-PH-Puro, Miaoling Biotechnology, Cat. No. P50541) using Lipofectamine 3000 (Invitrogen) for 48 hours. Subsequently, cells were treated or untreated with GDC-0941 in serum-free conditions for 24 hours and fixed with 4% paraformaldehyde. GFP signals were observed using a Leica DM6B Thunder fluorescence microscope to analyze the cellular distribution of PIP2.

[0130] (2) Experimental results

[0131] To evaluate whether GDC-0941 can induce methuosis in multiple cancer types, we tested its effect in a panel of cancer cell lines harboring genetic alterations in the PI3K pathway. E545K )、ME180(PIK3CA E545K ), MDA-MB-468 (PTEN deletion) and PANC-1 (Kras G12D ) showed obvious cytoplasmic vacuolation after GDC-0941 treatment under serum-free conditions. This phenotype can be +Blockade of this process by the ATPase inhibitor bafilomycin A1 confirmed that vacuolar dynamics play a role in this process. In contrast, PC-3 (PTEN deletion) and HCC70 (PTEN deletion) cells did not exhibit vacuolation under the same conditions. Immunofluorescence analysis further revealed that PIP2 underwent changes in vacuolar membrane localization in cell lines undergoing vacuolation. Treatment with the phospholipase C (PLC) agonist m-3M3FBS, which hydrolyzes PIP2, significantly inhibited vacuolation, suggesting that PIP2 plays a key role in methuosis-associated vacuolation. Notably, GDC-0941 induced significant cell death in CaSki (cervical cancer cell line), ME180 (cervical cancer cell line), MDA-MB-468 (breast cancer cell line), and PANC-1 (pancreatic cancer cell line), indicating that methuosis is an important mechanism of PI3K inhibition-induced cytotoxicity in these cells.

[0132] Macropinocytosis is an endocytic mechanism that is often activated under nutrient stress conditions and is a key prerequisite for the occurrence of methuosis. To evaluate macropinocytosis activity, we performed TMR-dextran uptake experiments under serum-free conditions. The results showed that after GDC-0941 treatment, CaSki, ME180, MDA-MB-468, PANC-1, OVSAHO, SNU119, and OVCAR4-PIK3CA E545K All cells showed sustained uptake of TMR-dextran, indicating that these cell lines maintained macropinocytosis after PI3K signaling inhibition. In contrast, HCC70 cells showed no significant dextran uptake, and PC-3 cells showed a significant decrease in uptake after GDC-0941 treatment, which may explain the lack of vacuolization in these two cell lines.

[0133] Taken together, these data indicate that GDC-0941-induced methuosis relies on two key factors: sustained PI3K-independent macropinocytic uptake and redistribution of PIP2 to the vacuole membrane, which synergistically drive GDC-0941-induced cytotoxic vacuolization.

[0134] In a specific embodiment, the present invention demonstrates that GDC-0941 acts synergistically with dietary restriction to induce methuosis in vivo, as shown in the experimental results. Figure 8 Specifically, as follows:

[0135] (1) Experimental methods

[0136] Eight-week-old female BALB / c nude mice were subcutaneously inoculated with 4×106 cells of ovarian cancer cells SNU119 mixed with Matrigel. 3 The animals were randomly divided into four groups: an ad libitum feeding group (AL), a restricted diet group (DR), a group treated with GDC-0941 under ad libitum feeding conditions (GDC+AL), and a group receiving GDC-0941 combined with a restricted diet (GDC+DR). The AL group was fed an irradiated purified rodent diet containing 3.601 kcal / g of physiological energy and consisting of 14.1% protein, 10% fat, and 72.7% carbohydrates. The DR group was fed 60% of the AL group's daily diet, supplemented with minerals and vitamins to prevent malnutrition. The diet maintained a similar texture and appearance to that of the AL group. GDC-0941 was dissolved in a 0.5% methylcellulose / 0.2% Tween-80 solution and administered orally by gavage at a dose of 120 mg / kg once daily for the entire treatment period. Tumor dimensions were measured with calipers every other day during the experiment, and tumor volume was calculated using the following formula: tumor volume = (length × width²) / 2.

[0137] Western blot analysis: Cells were lysed with ice-cold RIPA buffer containing protease and phosphatase inhibitors (Roche), and total protein was extracted. Proteins were separated by SDS-PAGE electrophoresis, transferred to a membrane, and incubated with primary antibodies overnight at 4°C. Images were analyzed using fluorescent secondary antibodies and the Odyssey infrared imaging system (Li-COR).

[0138] Histological Analysis: Tumor tissues were fixed overnight in 10% neutral-buffered formaldehyde and subsequently embedded in paraffin. Paraffin-embedded blocks were sectioned and stained with hematoxylin and eosin (H&E) for histological analysis. Stained sections were examined under a light microscope to assess their histopathological features.

[0139] (2) Experimental results

[0140] To evaluate the efficacy of PI3K inhibitor GDC-0941 combined with nutrient restriction to induce methoedosis in vivo, we established a mouse xenograft tumor model by subcutaneously injecting SNU119 cells (ovarian cancer cells). 3Mice were randomly divided into four treatment groups: ad libitum (AL), dietary restriction (DR), GDC-0941 treatment under ad libitum conditions (GDC+AL), and GDC-0941 combined with dietary restriction (GDC+DR). The AL group was given an irradiated purified rodent chow diet containing 3.601 kcal / g of energy and consisting of 14.1% protein, 10% fat, and 72.7% carbohydrates. The dietary restriction group was given a diet containing 60% of the calories of the ad libitum group daily, supplemented with minerals and vitamins to prevent malnutrition. The diet used was similar in texture and appearance to that of the control group. The GDC-0941 group, which was given ad libitum, was administered GDC-0941, dissolved in 0.5% methylcellulose / 0.2% Tween-80, by oral gavage at a dose of 120 mg / kg once daily for the entire treatment period. The treatment conditions for the GDC-0941 combined with dietary restriction group were to give the control group 60% of the calories of food daily and the same GDC-0941 treatment as above. Tumor growth assessment showed that dietary restriction alone slightly inhibited tumor growth, with a tumor growth inhibition rate (TGI) of 42%. In contrast, the use of GDC-0941 under free eating conditions significantly enhanced tumor regression (TGI of 68%, p<0.001). It is worth noting that the combination of GDC-0941 and dietary restriction (GDC+DR) achieved the most significant tumor inhibitory effect (TGI of 97%, p<0.001). Histological examination of tumor sections stained with hematoxylin-eosin (H&E) revealed obvious vacuolated cells, among which the combined treatment group had the strongest vacuolation effect. Furthermore, western blot analysis of tumor lysates revealed significant reductions in phosphorylated AKT (Ser473 and Thr308) and phosphorylated S6RP (Ser235 / Ser236) protein levels in the GDC+AL and GDC+DR treatment groups, confirming effective inhibition of the PI3K / AKT pathway. These in vivo results are consistent with our in vitro findings, indicating that GDC-0941, particularly when combined with dietary restriction, effectively induces methuosis and inhibits tumor growth. These data suggest that combining PI3K inhibition with dietary restriction is a promising anticancer therapeutic strategy.

[0141] Figure 2 Schematic diagram of an intelligent cancer diagnosis and treatment device based on the combination of the PI3K inhibitor GDC-0941 and dietary restriction, provided by an embodiment of the present invention. The device includes a memory and a processor; the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, it is used to perform the following method:

[0142] S101: Obtaining cancer diagnosis element information related to the patient;

[0143] S102: Determining whether the patient is a cancer patient based on the cancer diagnosis element data related to the patient;

[0144] S103: When the patient is a cancer patient, the following treatment regimen is selected: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient.

[0145] In some embodiments, the cancer diagnosis elements include medical history, symptom manifestations, examination data, imaging examination data, laboratory examination data and / or pathological examination data;

[0146] In some embodiments, if the examination data shows that a mass or tumor exists in the relevant tissue, the imaging examination data shows that a cystic or solid mass exists in the relevant tissue, the laboratory examination data shows that the CA125 value in the serum of the patient is higher than the normal range and / or the pathological examination data shows that the biopsy tissue is cancerous tissue, then the patient is judged to be a cancer patient.

[0147] In some embodiments, the intelligent cancer diagnosis and treatment device based on the combination of the PI3K inhibitor GDC-0941 and dietary restriction may further include an input device and an output device.

[0148] In some embodiments, the cancer is a cancer in which the PI3K signaling pathway is altered.

[0149] In some embodiments, the cancer is ovarian cancer, cervical cancer, breast cancer, or pancreatic cancer.

[0150] In some embodiments, the memory, processor, input device, and output device may be connected via a bus or other means. Figure 2 The bus connection mode is used as an example; the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, the above method is implemented.

[0151] In some embodiments, a memory may be understood as any device for storing a program, and a processor may be understood as a device for using a program.

[0152] In addition, an embodiment of the present invention further provides a computer-readable storage medium having a computer program stored thereon, wherein when the computer program is executed by a processor, the following method is implemented:

[0153] S101: Obtaining cancer diagnosis element information related to the patient;

[0154] S102: Determining whether the patient is a cancer patient based on the cancer diagnosis element data related to the patient;

[0155] S103: When the patient is a cancer patient, the following treatment regimen is selected: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient.

[0156] In some embodiments, the cancer diagnosis elements include medical history, symptom manifestations, examination data, imaging examination data, laboratory examination data and / or pathological examination data;

[0157] In some embodiments, if the examination data shows that a tumor or mass exists in the relevant tissue, the imaging examination data shows that a cystic or solid mass exists in the relevant tissue, the laboratory examination data shows that the CA125 value in the serum of the patient is higher than the normal range and / or the pathological examination data shows that the biopsy tissue is cancerous tissue, then the patient is judged to be a cancer patient.

[0158] In some embodiments, the cancer is a cancer in which the PI3K signaling pathway is altered.

[0159] In some embodiments, the cancer is ovarian cancer, cervical cancer, breast cancer, or pancreatic cancer.

[0160] In addition, an embodiment of the present invention further provides a method for inducing non-apoptotic death (methuosis) of cancer cells, comprising treating cancer cells with the PI3K inhibitor GDC-0941 in combination with dietary restriction.

[0161] In a specific embodiment of the present invention, the present invention experimentally demonstrates that the PI3K inhibitor GDC-0941 combined with dietary restriction can induce non-apoptotic cell death (methuosis) in cancer cells, and that the treatment regimen of the PI3K inhibitor GDC-0941 combined with dietary restriction has a synergistic therapeutic effect on the treatment of cancer (the specific experimental methods and experimental results are described above).

[0162] In some embodiments, the cancer cell is an ovarian cancer cell, a cervical cancer cell, a breast cancer cell, or a pancreatic cancer cell.

[0163] In some embodiments, the cancer cells include, but are not limited to, cancer cells derived from a subject and commercially available cancer cells.

[0164] In some embodiments, the commercially available cancer cells include but are not limited to: SNU119 cells, OVSAHO cells, OVCAR4 cells, CaSki cells, ME180 cells, PANC-1 cells, HCC70 cells, MDA-MB468 cells, and PC-3 cells.

[0165] In addition, an embodiment of the present invention further provides a system for inducing non-apoptotic death (methuosis) of cancer cells, the system comprising:

[0166] an acquisition unit, used for acquiring cancer cells to be processed;

[0167] The treatment unit treats the cancer cells with the PI3K inhibitor GDC-0941 in combination with dietary restriction to induce methuosis.

[0168] In some embodiments, the cancer cell is an ovarian cancer cell, a cervical cancer cell, a breast cancer cell, or a pancreatic cancer cell.

[0169] In addition, an embodiment of the present invention further provides a device for inducing non-apoptotic cell death (methuosis), the device comprising a memory and a processor; the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, it is used to perform the following method:

[0170] S101: Obtaining cancer cells to be processed;

[0171] S102: Treating the cancer cells with the PI3K inhibitor GDC-0941 in combination with dietary restriction to induce methuosis.

[0172] In some embodiments, the cancer cell is an ovarian cancer cell, a cervical cancer cell, a breast cancer cell, or a pancreatic cancer cell.

[0173] In addition, an embodiment of the present invention further provides a computer-readable storage medium having a computer program stored thereon, wherein when the computer program is executed by a processor, the following method is implemented:

[0174] S101: Obtaining cancer cells to be processed;

[0175] S102: Treating the cancer cells with the PI3K inhibitor GDC-0941 in combination with dietary restriction to induce methuosis.

[0176] In some embodiments, the cancer cell is an ovarian cancer cell, a cervical cancer cell, a breast cancer cell, or a pancreatic cancer cell.

[0177] In addition, an embodiment of the present invention further provides a method for treating cancer, comprising: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 in combination with dietary restriction to a subject in need thereof.

[0178] In some embodiments, the cancer is ovarian cancer, cervical cancer, breast cancer, or pancreatic cancer.

[0179] In some embodiments, the PI3K inhibitor GDC-0941 as described above can be administered to the subject by any one of oral administration, injection administration, local administration, or any combination thereof.

[0180] In some embodiments, the subject refers to any animal, including humans and non-human animals. Non-human animals include all vertebrates, for example, mammals, such as non-human primates (particularly higher primates), sheep, dogs, rodents (such as mice or rats), guinea pigs, goats, pigs, cats, rabbits, cows, and any livestock or pets; as well as non-mammals, such as chickens, amphibians, reptiles, etc. In a preferred embodiment, the subject is a human.

[0181] In some embodiments, the treatment refers to the medical management of a patient for the purpose of preventing, curing, ameliorating, or stabilizing a disease, pathological condition, or disorder. The term includes active therapy, i.e., treatment specifically for the purpose of ameliorating a disease, pathological condition, or disorder, and also includes causal therapy, i.e., treatment aimed at removing the cause of the disease, pathological condition, or disorder.

[0182] In some embodiments, the treatment also includes palliative treatment, which is treatment designed to relieve symptoms rather than cure the disease, pathological state or condition; the treatment also includes preventive treatment, which is treatment aimed at minimizing or partially or completely inhibiting the development of the relevant disease, pathological state or condition; and supportive treatment, which is treatment used to supplement another specific therapy for the purpose of improving the relevant disease, pathological state or condition.

[0183] Those skilled in the art will clearly understand that, for the convenience and brevity of description, the specific working processes of the systems, devices and units described above can refer to the corresponding processes in the aforementioned method embodiments and will not be repeated here.

[0184] In the several embodiments provided in this application, it should be understood that the disclosed systems, devices and methods can be implemented in other ways. For example, the device embodiments described above are merely schematic. For example, the division of the units is merely a logical function division. In actual implementation, there may be other division methods, such as multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed can be an indirect coupling or communication connection through some interfaces, devices or units, which can be electrical, mechanical or other forms.

[0185] The units described as separate components may or may not be physically separate, and the components shown as units may or may not be physical units, that is, they may be located in one place or distributed across multiple network units. Some or all of these units may be selected to achieve the purpose of this embodiment according to actual needs.

[0186] In addition, the functional units in the various embodiments of the present invention may be integrated into a single processing unit, each unit may exist physically separately, or two or more units may be integrated into a single unit. The aforementioned integrated units may be implemented in the form of hardware or software functional units.

[0187] Those skilled in the art will understand that all or part of the steps in the various methods of the above embodiments can be completed by instructing related hardware through a program, and the program can be stored in a computer-readable storage medium, which may include: read-only memory (ROM), random access memory (RAM), disk or optical disk, etc.

[0188] The above is a detailed introduction to a computer device provided by the present invention. For those skilled in the art, according to the concept of the embodiments of the present invention, there may be changes in the specific implementation methods and application scope. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. A cancer intelligent diagnosis and treatment system based on the combination of PI3K inhibitor GDC-0941 and dietary restriction, characterized by: The system comprises: A diagnostic element information collector is used to collect cancer diagnostic element information related to the patient and send the collected data results to the disease diagnosis matcher; A disease diagnosis matcher, used to match the diagnosis element information of the diagnosis element information collector with the disease type, and send the matched data results to the treatment plan matcher; A treatment plan matcher, configured to determine the matching result of the disease diagnosis matcher and call a treatment plan according to the matching result, and then send the treatment plan data result to the disease diagnosis and treatment information outputter; A disease diagnosis and treatment information output device, configured to output the data results received from the disease diagnosis matcher and the treatment plan matcher to a receiving unit; In the treatment plan matcher, if the disease diagnosis matcher matches the patient to be diagnosed as a cancer patient, the following treatment plan is called: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restrictions to the patient to be diagnosed; In the treatment plan matcher, if the disease diagnosis matcher matches the patient to be diagnosed as a non-cancer patient, then the following treatment plan is called without matching: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient to be diagnosed; Optionally, the cancer is ovarian cancer, cervical cancer, breast cancer or pancreatic cancer.

2. The cancer intelligent diagnosis and treatment system based on the combination of PI3K inhibitor GDC-0941 and dietary restriction according to claim 1, characterized in that: The cancer diagnosis elements include medical history, symptoms, examination data, imaging examination data, laboratory test data and / or pathological examination data; Optionally, the cancer is ovarian cancer, cervical cancer, breast cancer or pancreatic cancer.

3. The intelligent cancer diagnosis and treatment system based on the combination of PI3K inhibitor GDC-0941 and dietary restriction according to claim 2, characterized in that: In the disease diagnosis matching device, if the examination data shows the presence of a tumor or mass in the relevant tissue, the imaging examination data shows the presence of a cystic or solid mass, the laboratory examination data shows that the CA125 value in the patient's serum is higher than the normal range, and / or the pathological examination data shows that the biopsy tissue is cancerous tissue, the patient is matched as a cancer patient; In the disease diagnosis matching device, if the examination data shows that there is no abnormality in the surrounding tissue, the imaging examination data shows that there is no abnormality, the laboratory examination data shows that the CA125 value in the patient's serum is within the normal range and / or the pathological examination data shows that the biopsy tissue is normal tissue, the patient is matched as a non-cancer patient.

4. The cancer intelligent diagnosis and treatment system based on the combination of PI3K inhibitor GDC-0941 and dietary restriction according to claim 1, characterized in that: The combination of the PI3K inhibitor GDC-0941 and dietary restriction in the treatment regimen has a synergistic therapeutic effect on the treatment of cancer.

5. The cancer intelligent diagnosis and treatment system based on the combination of PI3K inhibitor GDC-0941 and dietary restriction according to claim 1, characterized in that: The receiving unit is a display screen, a computer client, a mobile phone client or a tablet.

6. An intelligent cancer diagnosis and treatment device based on the combination of PI3K inhibitor GDC-0941 and dietary restriction, characterized in that: The device includes a memory and a processor; the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, it is used to perform the following method: S101: Obtaining cancer diagnosis element information related to the patient; S102: Determining whether the patient is a cancer patient based on the cancer diagnosis element data related to the patient; S103: When the patient is a cancer patient, the following treatment regimen is selected: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient.

7. The intelligent cancer diagnosis and treatment device based on the combination of PI3K inhibitor GDC-0941 and dietary restriction according to claim 6, characterized in that: The cancer diagnosis elements include medical history, symptoms, examination data, imaging examination data, laboratory test data and / or pathological examination data; Optionally, if the examination data shows that a tumor or mass exists in the relevant tissue, the imaging examination data shows that a cystic or solid mass exists in the relevant tissue, the laboratory examination data shows that the CA125 value in the patient's serum is higher than the normal range, and / or the pathological examination data shows that the biopsy tissue is cancerous tissue, then the patient is determined to be a cancer patient; Optionally, the cancer is ovarian cancer, cervical cancer, breast cancer or pancreatic cancer.

8. A computer-readable storage medium having a computer program stored thereon, characterized in that: When the computer program is executed by a processor, the following method is implemented: S101: Obtaining cancer diagnosis element information related to the patient; S102: Determining whether the patient is a cancer patient based on the cancer diagnosis element data related to the patient; S103: When the patient is a cancer patient, selecting the following treatment plan: administering a therapeutically effective amount of the PI3K inhibitor GDC-0941 combined with dietary restriction to the patient; Optionally, the cancer is ovarian cancer, cervical cancer, breast cancer or pancreatic cancer.

9. A method for inducing non-apoptotic cell death (methuosis) of cancer cells, characterized in that: The method comprises treating cancer cells with the PI3K inhibitor GDC-0941 in combination with dietary restriction; Optionally, the cancer cells are ovarian cancer cells, cervical cancer cells, breast cancer cells or pancreatic cancer cells.

10. A system for inducing non-apoptotic cell death (methuosis) of cancer cells, characterized in that: The system comprises: an acquisition unit, used for acquiring cancer cells to be processed; a treatment unit, treating the cancer cells with the PI3K inhibitor GDC-0941 in combination with dietary restriction to induce methuosis; Optionally, the cancer cells are ovarian cancer cells, cervical cancer cells, breast cancer cells or pancreatic cancer cells.

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