Use of iron chelators in the treatment and / or prevention of endometrial cancer

Abstract

This invention discloses the application of iron chelators in the treatment and / or prevention of endometrial cancer. Iron chelators induce apoptosis in endometrial cancer cells and inhibit their growth. This invention also discovers that PTEN inhibitors can enhance the sensitivity of endometrial cancer to iron chelators. This invention provides new ideas and directions for the development of drugs for the treatment and / or prevention of endometrial cancer.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and relates to the application of iron chelators in treating and / or preventing endometrial cancer. Background Art

[0002] Endometrial cancer (EC) is an epithelial malignant tumor that occurs in the endometrium and is one of the three most common malignant tumors of the female reproductive tract. Treatment options for endometrial cancer include surgery, radiotherapy, chemotherapy, and other methods. Although the prognosis of patients with endometrial cancer has been significantly improved with the help of early diagnosis and continuous improvement of treatment methods, there are still many problems with EC drug treatment, such as drug resistance, toxic reactions, and poor efficacy. There is an urgent need to develop safer and more effective drugs to improve the efficacy and prognosis of EC and increase the survival rate of EC patients. Summary of the invention

[0003] In view of this, in order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide an iron chelator for use in the preparation of a drug for the treatment and / or prevention of endometrial cancer, wherein the iron chelator induces apoptosis of endometrial cancer cells or inhibits the tumor growth of endometrial cancer cells by chelating iron to achieve the effect of inhibiting the progression of endometrial cancer.

[0004] To achieve the above object, the present invention provides the following technical solution:

[0005] The first aspect of the present invention provides any of the following applications:

[0006] 1) Use of iron chelators in the preparation of drugs for the treatment and / or prevention of endometrial cancer;

[0007] 2) Use of iron chelators in the preparation of drugs for treating and / or preventing PTEN deficiency-related diseases;

[0008] 3) Application of PTEN inhibitors in enhancing the sensitivity of endometrial cancer to iron chelators.

[0009] Furthermore, the iron chelator is selected from one or more of deferoxamine, deferiprone and deferasirox.

[0010] Furthermore, the iron chelator is deferasirox.

[0011] Furthermore, the expression level of PTEN in endometrial cancer is decreased.

[0012] Furthermore, the PTEN deficiency-related diseases include cancer.

[0013] Furthermore, the cancer includes endometrial cancer, breast cancer, lung cancer, and glioma.

[0014] Furthermore, the cancer is endometrial cancer.

[0015] Furthermore, the PTEN inhibitors include nucleic acid inhibitors, protein inhibitors, and compounds.

[0016] Furthermore, the nucleic acid inhibitors include interfering RNA and antisense nucleic acids.

[0017] Furthermore, the interfering RNA includes shRNA, siRNA, dsRNA, and miRNA.

[0018] Furthermore, the interfering RNA is selected from shRNA.

[0019] Furthermore, the sequence of the shRNA is shown in SEQ ID NO.1-2.

[0020] A second aspect of the present invention provides a pharmaceutical composition for treating endometrial cancer, wherein the pharmaceutical composition comprises an iron chelator and / or a PTEN inhibitor.

[0021] Furthermore, the iron chelator is selected from one or more of deferoxamine, deferiprone and deferasirox.

[0022] Furthermore, the iron chelator is deferasirox.

[0023] Furthermore, the PTEN inhibitors include nucleic acid inhibitors, protein inhibitors, and compounds.

[0024] Furthermore, the nucleic acid inhibitors include interfering RNA and antisense nucleic acid.

[0025] Furthermore, the interfering RNA includes shRNA, siRNA, dsRNA, and miRNA.

[0026] Furthermore, the interfering RNA is selected from shRNA.

[0027] Furthermore, the sequence of the shRNA is shown in SEQ ID NO.1-2.

[0028] Furthermore, the pharmaceutical composition includes a pharmaceutically acceptable carrier and / or excipient.

[0029] Furthermore, the pharmaceutically acceptable carrier and / or excipient includes a diluent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant and / or a disintegrant.

[0030] Furthermore, the pharmaceutical composition may be administered in any convenient pharmaceutical dosage form.

[0031] The third aspect of the present invention provides a drug comprising a PTEN inhibitor.

[0032] Furthermore, the PTEN inhibitors include nucleic acid inhibitors, protein inhibitors, and compounds.

[0033] Furthermore, the nucleic acid inhibitors include interfering RNA and antisense nucleic acid.

[0034] Furthermore, the interfering RNA includes shRNA, siRNA, dsRNA, and miRNA.

[0035] Furthermore, the nucleic acid inhibitor is selected from shRNA.

[0036] Furthermore, the sequence of the shRNA is shown in SEQ ID NO.1-2.

[0037] The fourth aspect of the present invention provides a method for inducing apoptosis of endometrial cancer cells or inhibiting the growth of endometrial cancer cells in vitro, the method comprising administering the pharmaceutical composition according to the second aspect of the present invention.

[0038] Further, the steps of inducing apoptosis of endometrial cancer cells in vitro or inhibiting the growth of endometrial cancer cells in vitro are as follows:

[0039] (1) Treating endometrial cancer cells with the pharmaceutical composition provided by the present invention;

[0040] (2) Observe the apoptosis or growth of endometrial cancer cells.

[0041] Furthermore, the endometrial cancer cells include Ishikawa, AN3-CA, HEC-1-B, RL95-2, HEC-1-A, MFE-280, MFE-296, SNG-M, HECCL-1, KLE, ECC-1, ECC-10, ECC-12, HEC-151, HEC-251, and SK-UT-1.

[0042] Furthermore, the endometrial cancer cells are selected from Ishikawa and AN3-CA.

[0043] The fifth aspect of the present invention provides use of the pharmaceutical composition according to the second aspect of the present invention in inducing apoptosis of endometrial cancer cells or inhibiting the growth of endometrial cancer cells in vitro.

[0044] Furthermore, the endometrial cancer cells are selected from one or more of Ishikawa, AN3-CA, HEC-1-B, RL95-2, HEC-1-A, MFE-280, MFE-296, SNG-M, HECCL-1, KLE, ECC-1, ECC-10, ECC-12, HEC-151, HEC-251, and SK-UT-1.

[0045] Furthermore, the endometrial cancer cells are selected from Ishikawa and AN3-CA.

[0046] The present invention has the following advantages and beneficial effects:

[0047] The present invention provides the use of an iron chelator in preparing a drug for treating and / or preventing endometrial cancer, which is disclosed for the first time. The present invention finds that the iron chelator can induce apoptosis of endometrial cancer cells and inhibit tumor growth of endometrial cancer cells. The present invention also finds that reducing the expression level of PTEN can enhance the sensitivity of endometrial cancer to the iron chelator, which provides a new method and new idea for the clinical treatment of endometrial cancer, and has significant significance in the adjuvant treatment and prevention and treatment of endometrial cancer. Therefore, the iron chelator provided by the present invention has very good clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 The figures show the enhanced sensitivity of endometrial cancer cells to deferasirox (DFX) with PTEN knockdown, among which 1A shows the enhanced sensitivity of endometrial cancer cells to deferasirox (DFX) with PTEN deletion; 1B shows the enhanced sensitivity of KLE cells to deferasirox (DFX) after PTEN knockdown; 1C shows the enhanced sensitivity of HEC-1-B cells to deferasirox (DFX) after PTEN knockdown; 1D shows the reduced sensitivity of RL95-2 cells overexpressing PTEN to deferasirox (DFX).

[0049] Figure 2 These are the experimental results of deferasirox (DFX) inducing apoptosis of PTEN-deficient endometrial cancer cells, where 2A is the result of AN3-CA cell apoptosis detection after the cells were treated with 50 μM deferasirox (DFX) for 48 hours, where the left figure is the flow cytometry result, and the right figure is the statistical result of the early and late apoptotic cell ratios; 2B is the result of cell apoptosis detection after ISHIKAWA cells were treated with 50 μM deferasirox (DFX) for 48 hours, where the left figure is the flow cytometry result, and the right figure is the statistical result of the early and late apoptotic cell ratios.

[0050] Figure 3 Figures 3A, 3B, and 3C are experimental results of Deferasirox (DFX) inhibiting tumor growth in PTEN-deficient endometrial cancer cells; Figure 3D is an experimental result of the effect of Deferasirox (DFX) on the body weight of nude mice; Figure 3E is an experimental result of immunohistochemistry to detect the effect of Deferasirox (DFX) on proliferation indicators (Ki67) and apoptosis indicators (Cleaved-caspase3). DETAILED DESCRIPTION

[0051] The following provides definitions of some terms used in this specification. Unless otherwise defined, all technical and scientific terms used in the present invention generally have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs.

[0052] The present invention provides the use of an iron chelator in preparing a medicine for treating and / or preventing endometrial cancer.

[0053] In the present invention, the term "iron chelator" refers to a compound that can form a complex with iron ions. The iron chelator can be selected from one or more of deferoxamine, deferiprone, and deferasirox. In a specific embodiment of the present invention, the iron chelator is deferasirox.

[0054] In the present invention, the term "treatment and / or prevention" may refer to therapeutic treatment or preventive measures, wherein the goal is to prevent or slow down (mitigate) an undesirable physiological condition, disorder or disease, or to obtain a beneficial or desired clinical result. Beneficial or desired clinical results include, but are not limited to, relieving symptoms; alleviating the extent of a condition, disorder or disease; stabilizing (i.e., not worsening) the state of a condition, disorder or disease; delaying the onset of a condition, disorder or disease or slowing its progression; improving a condition, disorder or disease state; and alleviating (whether partial or complete) (whether detectable or undetectable) or improving or ameliorating a condition, disorder or disease. Treatment may include causing a clinically significant response without excessive side effects. Treatment also includes a prolonged survival period compared to the expected survival period when not receiving treatment.

[0055] The present invention provides the use of an iron chelator in preparing a medicine for treating and / or preventing PTEN deficiency-related diseases.

[0056] In the present invention, the term "PTEN" is a tumor suppressor gene. Due to its phosphatase activity, PTEN protein can modify other proteins by removing phosphate groups, thereby preventing cells from growing and dividing too quickly or dividing uncontrollably, causing cells to stop dividing and enter apoptosis, thereby achieving regulation of the cell division cycle and inhibiting tumor formation. PTEN loss leads to malignant tumor progression and drug resistance.

[0057] In the present invention, PTEN deficiency may refer to a reduction or loss of PTEN expression level, or a reduction or loss of PTEN activity function.

[0058] In the present invention, PTEN deficiency-related diseases include cancer, wherein cancer includes but is not limited to endometrial cancer, colorectal cancer, pancreatic cancer, hepatocellular carcinoma, bile duct cancer, gallbladder cancer, ovarian cancer, breast cancer, lung cancer, esophageal cancer, gastric cancer, renal cell carcinoma, ureteral cancer, bladder cancer, prostate cancer, cervical cancer, glioma. In one embodiment, cancer is selected from endometrial cancer, breast cancer, lung cancer, glioma. In a specific embodiment of the present invention, the PTEN deficiency-related disease is endometrial cancer.

[0059] The present invention provides the use of a PTEN inhibitor in enhancing the sensitivity of endometrial cancer to an iron chelator.

[0060] In the present invention, the term "inhibitor" refers to any substance that can inhibit the activity of PTEN protein, inhibit the stability of PTEN gene or protein, inhibit the expression level of PTEN, inhibit the effective action time of PTEN protein, and inhibit the activity of PTEN.

[0061] Wherein, the inhibitor includes nucleic acid inhibitor, protein inhibitor, compound or combination thereof. As one embodiment of the present invention, the "inhibitor" is a substance that inhibits the expression level of PTEN.

[0062] Among them, the nucleic acid inhibitor is selected from: interfering molecules that use PTEN or its transcript as the target sequence and can inhibit PTEN gene expression or gene transcription, including but not limited to shRNA, siRNA, ribozymes, antisense nucleic acids, dsRNA, microRNA, zinc fingers, CRISPR / Cas9, or constructs that can express or form the shRNA, siRNA, ribozymes, antisense nucleic acids, dsRNA, microRNA, zinc fingers, CRISPR / Cas9.

[0063] Among them, siRNA can include partially purified RNA, substantially pure RNA, synthetic RNA or recombinantly produced RNA and RNA that is different from the altered natural RNA by adding, deleting, replacing and / or changing one or more nucleotides. The alteration can include adding non-nucleotide substances, such as adding to the end of the siRNA or one or more internal nucleotides of the siRNA; making the siRNA resistant to ribozyme digestion (e.g., using 2'-substituted ribonucleotides or modifying the sugar phosphate backbone); or replacing one or more nucleotides in the siRNA with deoxyribonucleotides.

[0064] Among them, shRNA is a non-coding small RNA molecule that can form a hairpin structure. shRNA can inhibit gene expression through the RNA interference pathway. When shRNA is introduced into cells, it will be recognized by the intracellular ribozyme Dicer and cut into small interfering RNA (siRNA) of about 21 nucleotides. siRNA binds to a protein complex called RNA-induced silencing complex (RISC). The Argonaute protein in RISC uses the antisense strand of siRNA to recognize and bind to the target mRNA, and then degrades the target mRNA by cutting or inhibiting translation, thereby achieving the inhibition of specific gene expression. In one embodiment of the present invention, the nucleic acid inhibitor is selected from shRNA. In a specific embodiment of the present invention, the shRNA sequence is shown in SEQ ID NO.1-2.

[0065] Wherein, the protein inhibitor is selected from substances capable of inhibiting PTEN protein, and the protein inhibitor includes proteolytic enzymes and protein binding molecules. The proteolytic enzyme is selected from enzymes capable of catalyzing the hydrolysis of PTEN protein. The protein binding molecule is selected from substances that specifically bind to PTEN protein, such as antibodies or ligands capable of inhibiting the activity of PTEN protein.

[0066] The present invention provides a pharmaceutical composition for treating endometrial cancer, wherein the pharmaceutical composition comprises the iron chelator and / or PTEN inhibitor as described above.

[0067] In the present invention, the term "pharmaceutical composition" refers to a composition comprising at least one biologically active compound. The iron chelator and the PTEN inhibitor may be selected from the iron chelators and PTEN inhibitors described above. In a specific embodiment of the present invention, the pharmaceutical composition comprises both an iron chelator and a PTEN inhibitor, wherein the iron chelator is deferasirox, and the PTEN inhibitor is shRNA, and the sequence of the shRNA is shown in SEQ ID NO.1-2.

[0068] The pharmaceutically acceptable carrier and / or adjuvant of the present invention include any material suitable for humans and / or mammals without excessive adverse side effects (such as toxicity, irritation and allergic reactions), that is, a material with a reasonable benefit / risk ratio. The pharmaceutically acceptable carrier and / or adjuvant are used as needed to help the stability of the formulation or to help improve its activity or its biological effectiveness or to produce an acceptable taste or smell in the case of oral administration. The pharmaceutical composition thus prepared can be administered by any appropriate administration method known to those skilled in the art as needed. When using the pharmaceutical composition, the pharmaceutical composition of the present invention is administered to humans at a safe and suitable dosage.

[0069] The pharmaceutical composition of the present invention may be contained in a pharmaceutically acceptable carrier and / or excipient, wherein the pharmaceutically acceptable carrier and / or excipient includes a diluent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant and / or a disintegrant. Among them, diluents include but are not limited to lactose, sodium chloride, glucose, urea, starch, and water; adhesives include but are not limited to starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methyl cellulose, carboxymethyl cellulose, ethyl cellulose, polyvinyl alcohol, polyethylene glycol, polyvinyl pyrrolidone, alginic acid and alginate, xanthan gum, hydroxypropyl cellulose and hydroxypropyl methylcellulose; surfactants include but are not limited to polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, monoglyceride of stearic acid, and hexadecanol; humectants include but are not limited to glycerol and starch; adsorption carriers include but are not limited to starch, lactose, bentonite, silica gel, kaolin, and bentonite; lubricants include but are not limited to zinc stearate, glyceride monostearate, polyethylene glycol, talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearyl fumarate, polyoxyethylene monostearate, monolauric sucrose acid ester, sodium lauryl sulfate, magnesium lauryl sulfate, and magnesium lauryl sulfate.

[0070] The suitable dosage of the pharmaceutical composition of the present invention can be prescribed in a variety of ways depending on factors such as the formulation method, administration method, patient's age, weight, gender, morbidity, diet, administration time, administration route, excretion rate and reaction sensitivity. A skilled physician can usually easily determine the prescription and the desired dosage that is effective for treatment.

[0071] The pharmaceutical composition of the present invention can be administered in any convenient pharmaceutical dosage form, including parenteral dosage forms and enteral dosage forms.

[0072] Among them, the non-gastrointestinal dosage forms include injection dosage forms, respiratory tract dosage forms, cavity dosage forms, mucosal dosage forms, and skin dosage forms; the gastrointestinal dosage forms include tablets, granules, capsules, solutions, dry suspensions, powders, sustained-release agents, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

[0073] The injectable dosage forms include but are not limited to intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections and intracavitary injections; the respiratory tract dosage forms include but are not limited to sprays, aerosols, and powder aerosols; the cavity administration dosage forms include but are not limited to suppositories, aerosols, effervescent tablets, drops, and pills, which can be used for including but not limited to the rectum, vagina, urethra, nasal cavity, and ear canal; the mucosal administration dosage forms include but are not limited to eye drops, nasal drops, eye ointments, gargles, sublingual tablets, adhesive tablets, and patches; the skin administration dosage forms include but are not limited to external solutions, lotions, liniments, ointments, plasters, pastes, and patches.

[0074] The iron chelator and PTEN inhibitor in the pharmaceutical composition of the present invention can be administered in any suitable manner, including simultaneous administration or sequential administration. When administered sequentially, the administration methods include: administering the iron chelator first and then the PTEN inhibitor; administering the PTEN inhibitor first and then the iron chelator.

[0075] The pharmaceutical composition of the present invention can also be used in combination with other drugs for treating endometrial cancer, and when used in combination, the drugs can be administered in any suitable manner, including simultaneous administration or sequential administration.

[0076] The present invention provides a method for inducing apoptosis of endometrial cancer cells or inhibiting the growth of endometrial cancer cells in vitro, the method comprising administering the pharmaceutical composition of the present invention as described above. In a specific embodiment of the present invention, the method comprises treating endometrial cancer cells with the pharmaceutical composition provided by the present invention; and observing the apoptosis or growth of endometrial cancer cells.

[0077] Among them, the endometrial cancer cells include Ishikawa, AN3-CA, HEC-1-B, RL95-2, HEC-1-A, MFE-280, MFE-296, SNG-M, HECCL-1, KLE, ECC-1, ECC-10, ECC-12, HEC-151, HEC-251, and SK-UT-1. In a specific embodiment of the present invention, the endometrial cancer cells are Ishikawa and AN3-CA.

[0078] The present invention will be further described in detail below in conjunction with the examples. The following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0079] Example 1 PTEN knockdown enhances the sensitivity of endometrial cancer cells to deferasirox (DFX)

[0080] 1. Experimental Materials

[0081] Anti-GAPDH was purchased from Abotek Co., Ltd., CCK8 (Yisheng Biotechnology Co., Ltd.), trypan blue (Beijing Zhongshan Biotechnology Co., Ltd.), and the lentiviral-mediated shRNA vector targeting human PTEN and the lentiviral plasmid for overexpression of human PTEN were purchased from Jikai Biotechnology Co., Ltd. (Shanghai, China).

[0082] The sources and culture conditions of endometrial cancer cell lines are shown in Table 1:

[0083] Table 1 Endometrial cancer cell lines

[0084]

[0085] 2. Experimental methods

[0086] 1) Construction of PTEN knockdown or overexpression endometrial cancer cell lines

[0087] Transduce cells at 30% confluency with 20-40 mol of lentivirus mixed with 40 μL of HiTransGA. After transduction, puromycin selection was applied to establish stably transfected cell lines. Successful gene disruption or overexpression was verified by immunoblotting for the corresponding target protein. The shRNA sequences used are shown in Table 2.

[0088] Table 2 shRNA sequences targeting PTEN

[0089]

[0090] 2) Western blot detection of PTEN protein expression in cells with knockdown or overexpression of PTEN

[0091] Expanded cultured cells were lysed with RIPA lysis buffer and protein was quantified by BCA method. Proteins were separated on NuPAGE 10% Bis-Tris gel and transferred to nitrocellulose membrane (Merck Millipore, Darmstadt, Germany). The membrane was blocked for 1 hour and 10 minutes and then incubated with primary antibody overnight at 4°C. After washing three times with TBST (10 mM Tris-HCl (pH 8), 150 mM NaCl, 0.05% Tween20), the membrane was incubated with secondary antibody for 2 hours at room temperature. After the final three washes with TBST, the membrane was developed using LI-COR Odyssey infrared scanner.

[0092] 3) Cell proliferation assay (CCK8)

[0093] Endometrial cancer cell lines, endometrial cancer cell lines with knockdown or overexpression of PTEN were digested to obtain cell suspensions. After counting the cells with trypan blue, the dilution method of the cell suspension was calculated so that the number of cells in each well of the 96-well plate reached a density of 5000-9000 / 200 μL, and the cells were continued to be cultured in an incubator; drug addition: after the cells adhered to the wall, drugs (deferasirox, DFX) were added for treatment; detection: according to the required experimental conditions, 20 μL of CCK8 reagent was added to each well after 48 hours, and the incubation continued for 2-4 hours, the OD450 was measured, and the cell proliferation was calculated based on the OD450 results.

[0094] 3. Experimental results

[0095] Five endometrial cancer cell lines were selected, and Western Blot results showed that PTEN expression was lost in AN3-CA, ISHIKAWA and RL95-2 cells, and PTEN was normally expressed in KLE and HEC-1-B cells. Cell proliferation assay results showed that PTEN-deficient endometrial cancer cells AN3-CA, ISHIKAWA and RL95-2 were more sensitive to deferasirox (DFX). Figure 1 A).

[0096] Western Blot results showed that compared with PTEN wild-type (shPTEN-NC) cells, the PTEN expression level in the PTEN knockout group was reduced in KLE and HEC-1-B cells. Cell proliferation assay results showed that knocking down PTEN enhanced the sensitivity of KLE and HEC-1-B cells to deferasirox (DFX). Figure 1 BC).

[0097] Western Blot results showed that compared with PTEN-deficient endometrial cancer cells RL95-2, the PTEN-overexpressing group expressed PTEN. Cell proliferation assay results showed that overexpression of PTEN reduced the sensitivity of RL95-2 cells to deferasirox (DFX). Figure 1 D).

[0098] Note: *(p<0.05), **(p<0.01), ***(p<0.001), ****(p<0.0001)

[0099] Example 2 Deferasirox (DFX) induces apoptosis of PTEN-deficient endometrial cancer cells

[0100] 1. Experimental Materials

[0101] Annexin V-647 and propidium iodide (PI) detection kit were purchased from Yisheng Biotechnology Co., Ltd. (Shanghai, China).

[0102] 2. Experimental methods

[0103] ISHIKAWA and AN3-CA cells were treated with DMSO or 50 μM DFX for 24 h. Cells were stained using Annexin V-647 and propidium iodide (PI) detection kit (Yeasen, Shanghai, China). Cells were harvested, washed with PBS, and resuspended in 1× binding buffer. 100 μL aliquots of the cell suspension were transferred to 5 mL culture tubes, and then 5 μL of Annexin V-647 and 10 μL of PI were added. The samples were incubated at room temperature in the dark for 15 min. After adding 200 μL of binding buffer to each tube, apoptotic cells were quantified using an AccuriC6 flow cytometer (BD Biosciences, East Rutherford, NJ, USA).

[0104] 3. Experimental results

[0105] After the cells were treated with 50 μM deferasirox (DFX) for 48 hours, the apoptotic cell ratio of AN3-CA and ISHIKAWA cells was determined by flow cytometry. Compared with the control cells, the proportion of early apoptotic cells in AN3-CA treated with deferasirox (DFX) increased from 1.72% to 6.7%, and the proportion of late apoptotic cells increased from 1.69% to 4.34% ( Figure 2 A). The proportion of early apoptotic cells in ISHIKAWA treated with Deferasirox (DFX) increased from 6.07% to 30.5%, and the proportion of late apoptotic cells increased from 3.5% to 11.1% ( Figure 2 B).

[0106] Note: *(p<0.05), **(p<0.01), ***(p<0.001), ****(p<0.0001)

[0107] Example 3 Deferasirox (DFX) inhibits tumor growth of PTEN-deficient endometrial cancer cells

[0108] 1. Experimental Materials

[0109] Deferasirox (DFX) was purchased from Novartis (Basel, Switzerland). For in vivo studies, DFX was dissolved in sodium chloride solution (0.9% w / v; CSPC Pharmaceutical Group. Shijiazhuang, Hebei, China).

[0110] 2. Experimental methods

[0111] 1) Nude mouse tumor formation experiment

[0112] All animal studies were performed in accordance with protocols approved by the Animal Center of the Institute of Basic Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College (Ethics Code: ACUC-A01-2023-039). Female BALB / c nude mice (4–6 weeks old) were purchased from HFKBio-Technology (Beijing, China). 6 A suspension of 100 μL of AN3-CA cells in 50% Matrigel (#354234, BD Biosciences, San Jose, CA, USA) was subcutaneously injected into the right dorsal flank of each mouse. Once the tumor volume reached 50–100 mm 3 The mice were randomly divided into groups. The treatment group was intraperitoneally injected with 300 mg / kg of deferasirox (DFX) every other day for 15 days, and the control group was injected with an equal volume of saline. Tumor volume and body weight were recorded every other day, and tumor volume was calculated using the following formula: Volume = length × width 2 / 2.

[0113] 2) Histology and immunohistochemistry (IHC)

[0114] Tissues were fixed in paraformaldehyde. H&E staining, DAB-enhanced Prussian blue staining, and IHC were performed by Servicebio (Wuhan, Hubei, China). Sections were examined using a PannoramicDESK microscope and analyzed using CaseViewer software (CV version 2.3).

[0115] 3. Experimental results

[0116] 1) Deferasirox (DFX) treatment inhibited tumor growth in AN3-CA cells ( Figure 3 A), resulting in a decrease in tumor volume and weight ( Figure 3 BC), the effect on the body weight of nude mice was negligible ( Figure 3 D).

[0117] 2) Immunohistochemical staining showed that treatment with Deferasirox (DFX) reduced the expression of proliferation markers (Ki67) and increased the expression of apoptosis markers (Cleaved-caspase3) ( Figure 3 E).

[0118] Note: *(p<0.05), **(p<0.01), ***(p<0.001), ****(p<0.0001)

[0119] The description of the above embodiments is only used to understand the method and core idea of ​​the present invention. It should be pointed out that, for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principle of the present invention, and these improvements and modifications will also fall within the scope of protection of the claims of the present invention.

Claims

1. Use of a PTEN inhibitor in the preparation of a drug for enhancing the sensitivity of endometrial cancer to iron chelators, characterized in that: The iron chelator is deferasirox, and the PTEN inhibitor is an interfering RNA having a sequence as shown in SEQ ID NO.1-2.

2. A pharmaceutical composition for treating endometrial cancer, characterized in that: The pharmaceutical composition comprises deferasirox and a PTEN inhibitor, wherein the PTEN inhibitor is an interfering RNA with a sequence as shown in SEQ ID NO.1-2.

3. The pharmaceutical composition according to claim 2, characterized in that The pharmaceutical composition includes a pharmaceutically acceptable carrier and / or excipient.

Images