Cell radiosensitivity determination method

Abstract

One or more embodiments of the invention provide a cell radiosensitivity determination method, which comprises the following steps: respectively processing a plurality of cell test groups of target cells by using a preset cell cycle arrest method to obtain a plurality of cell test groups with different G1 phase proportions; wherein the G1 phase proportion is the proportion of cells in the G1 phase in the cell test group; performing radiation operation on the plurality of cell test groups, and determining cell biological indexes in each cell test group after the radiation operation; and on the basis of the G1 phase proportion of each cell test group and the corresponding cell biological index, constructing a data model for indicating the corresponding relationship between the G1 phase proportion of the target cell and the cell biological index, and evaluating the radiation sensitivity of the target cell.

Description

Technical Field

[0001] One or more embodiments of the present specification relate to the field of biotechnology, and more particularly to a method for determining cell radiation sensitivity. Background Art

[0002] The radiation response of cells is usually closely related to the cell cycle they are in, and there are significant differences in the sensitivity to radiation at different cell cycle stages. Therefore, exploring the relationship between the cell cycle and the survival rate of tumor cells after radiation is of great research value.

[0003] Related studies mainly use methods such as clone formation experiments, heavy ion irradiation based on cell cycle synchronization, linear quadratic (LQ) model, flow cytometry, etc. However, these technologies have their own limitations, such as long experimental cycles, complex technical operations, insufficient sensitivity, or inability to comprehensively consider factors such as cell microenvironment, and there is still a lack of in-depth quantitative research on the relationship between cell cycle and cell radiation sensitivity. Summary of the invention

[0004] In view of this, one or more embodiments of this specification provide the following technical solutions:

[0005] According to a first aspect of one or more embodiments of this specification, a method for determining cell radiation sensitivity is provided, comprising:

[0006] A preset cell cycle arrest method is used to process multiple cell test groups of target cells respectively, so as to obtain multiple cell test groups with different G1 phase proportions; wherein the G1 phase proportion is used to indicate the proportion of cells in the G1 phase in the cell test group;

[0007] Performing radiation operation on the multiple cell test groups, and determining cell biological indicators in each cell test group after the radiation operation;

[0008] Based on the G1 phase proportions and corresponding cell biological indicators of each cell test group, a data model for indicating the corresponding relationship between the G1 phase proportions of the target cells and the cell biological indicators is constructed to evaluate the radiation sensitivity of the target cells.

[0009] Furthermore, the cell biological index includes cell survival rate.

[0010] Furthermore, the corresponding relationship between the G1 phase proportion of the target cells and the cell biological index includes: a relationship curve between the G1 phase proportion of the target cells and the cell survival rate.

[0011] Furthermore, the method of using a preset cell cycle arrest method to process multiple cell test groups of target cells respectively includes:

[0012] Adding doxycycline Dox to multiple cell test groups of target cells to induce the expression level of retinoblastoma tumor suppressor protein Rb protein in the target cells under Dox treatment;

[0013] By adjusting the Dox concentration and / or treatment time in each cell test group, the G1 phase proportion was adjusted with different Rb protein expression levels.

[0014] Furthermore, the method further comprises:

[0015] Based on the Dox concentration and the corresponding Rb protein expression level in each cell test group, the corresponding relationship between the Dox concentration and the Rb protein expression level was constructed.

[0016] Furthermore, the method further comprises:

[0017] Based on the Rb protein concentration and the corresponding G1 phase proportion of each cell test group, the corresponding relationship between the Rb protein concentration and the G1 phase proportion of the target cell is constructed.

[0018] Further, performing radiation operation on the multiple cell test groups includes:

[0019] Each test group of cells was exposed to a different dose of radiation.

[0020] Furthermore, the data model is also used to indicate the corresponding relationship between the cell cycle and the cell survival rate of the target cells under different radiation doses.

[0021] Furthermore, the target cells are tumor cells.

[0022] Furthermore, the tumor cells are head and neck tumor cell lines.

[0023] It can be seen from the above embodiments that the present specification can obtain multiple cell test groups with different G1 phase proportions by performing cell cycle arrest treatment on the target cells, and then perform radiation operation on each cell test group respectively, and record the cell biological indicators after the radiation operation, so as to quantitatively analyze the relationship between the cell cycle and the cell radiation sensitivity, and by constructing the corresponding data model, provide a reliable and accurate theoretical basis and data support for subsequent research in the fields of cell biology, radiation biology, and personalized treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a flow chart of a method for determining cell radiation sensitivity provided by an exemplary embodiment.

[0025] Figure 2 It is a flowchart of a method for determining cell radiation sensitivity provided by an exemplary embodiment.

[0026] Figure 3 It is a block diagram of a device for determining cell radiation sensitivity provided by an exemplary embodiment. DETAILED DESCRIPTION

[0027] The user information (including but not limited to user device information, user personal information, etc.) and data (including but not limited to data used for analysis, stored data, displayed data, etc.) involved in this manual are all information and data authorized by the user or fully authorized by all parties, and the collection, use and processing of relevant data must comply with the relevant laws, regulations and standards of relevant countries and regions, and provide corresponding operation entrances for users to choose to authorize or refuse.

[0028] The radiation response of cells is usually closely related to the cell cycle they are in, and there are significant differences in the sensitivity to radiation at different cell cycle stages. Therefore, exploring the relationship between the cell cycle and the survival rate of tumor cells after radiation is of great research value.

[0029] The relevant technologies mainly adopt the following methods:

[0030] Clone formation experiment: By directly measuring the activity of cells after radiation, a survival curve is drawn to obtain radiation biological parameters. However, this method has the problem of long experimental cycle. It usually takes several days to several weeks to observe cell survival, resulting in delayed data acquisition. In addition, the sensitivity of the measurement is low at low doses of radiation.

[0031] Heavy ion irradiation based on cell cycle synchronization: Modeling cell dynamics through experimental data and tracking the dynamic changes of cells under different irradiation protocols. The disadvantage of this method is that stress response may occur during cell cycle synchronization, changing the radiation sensitivity of cells, and it is difficult to ensure consistency in synchronization effects between different experiments, affecting the repeatability and reliability of the data.

[0032] Linear Quadratic (LQ) model: quantifies cell radiation sensitivity through parameters α and β, and fits the cell survival curve. However, the LQ model assumes that the radiation response has linear and quadratic effects, which may not accurately reflect the actual biological response in complex situations such as high doses or split doses. In addition, the model requires a large amount of data support and cannot explain individual differences.

[0033] Flow cytometry: used to accurately measure cell cycle distribution, especially to mark S-phase cells, so as to analyze the response of cell populations to radiation in different cycles. Although flow cytometry can provide these data, its operation is complicated, the result interpretation needs to combine multiple markers, and it mainly analyzes the transient state of cells and cannot provide long-term survival information.

[0034] In summary, the above technologies have their own limitations, such as long experimental cycle, complex operation, insufficient sensitivity or difficulty in comprehensively considering factors such as the cell microenvironment. These shortcomings affect the accurate assessment of cell radiation sensitivity.

[0035] The cell cycle can be divided into two major parts: interphase and mitosis (M). Interphase is divided into three subphases: Gap 1 (G1), DNA synthesis (S), and Gap 2 (G2). When facing radiation, G1 phase cells are usually more tolerant, S phase cells have relatively low radiation sensitivity, and G2 and M phase cells are the most sensitive to radiation.

[0036] In view of this, this specification proposes a method for determining cell radiation sensitivity. By adjusting the proportion of the pre-interphase G1 phase of the target cells and combining the cell biological indicators after radiation operation, the influence of the proportion of the G1 phase on the radiation sensitivity of the target cells is determined, and a data model is constructed to evaluate the radiation sensitivity of the target cells.

[0037] In order to enable those skilled in the art to better understand the technical solutions in the present application, the technical solutions in this specification will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application.

[0038] See also Figure 1 , Figure 1 An exemplary embodiment provides a method for determining cell radiation sensitivity. The method may include the following steps.

[0039] S101. Using a preset cell cycle arrest method, respectively process multiple cell test groups of target cells to obtain multiple cell test groups with different G1 phase proportions; wherein the G1 phase proportion is used to indicate the proportion of cells in the G1 phase in the cell test group.

[0040] In one embodiment, first, multiple cell test groups can be prefabricated. For example, target cells are cultured under preset culture conditions (such as 37°C, 5% CO2, 10% fetal bovine serum, etc.) to obtain multiple cell test groups. In order to ensure the stability and repeatability of the experimental results, the pre-prepared cell test group should ensure that the target cells are in the logarithmic growth phase. The cell division rate and proliferation rate at this stage are relatively constant, the distribution of each cell cycle stage is balanced, and it is not easily affected by external environmental fluctuations. Therefore, ensuring that the cells are in the logarithmic growth phase not only contributes to the stability of the cell cycle, but also improves the reliability and consistency of the experimental data, reduces errors, and ensures that the cell's response to external stimuli (such as radiation, drugs, etc.) has high reliability.

[0041] Next, each cell test group is processed separately using a preset cell cycle arrest method, and the G1 phase proportion of the target cells in each cell test group is adjusted to obtain a series of cell test groups with different G1 phase proportions.

[0042] There are many methods for cell cycle arrest, such as drug-induced arrest, temperature-induced arrest, and RNA interference. Drug-induced arrest: For example, drugs such as cytarabine (Ara-C), mimosine, and aphidicolin are used to specifically arrest cells in the G1 phase or S phase.

[0043] The G1 phase proportions of the multiple cell test groups obtained after adjustment can be randomly distributed or precisely controlled within a predetermined interval, such as 30%-40%, 40%-50%, etc. It can also be further controlled within multiple specified target G1 phase proportions, such as 20%, 30%, 40%... Among them, the target G1 phase proportion can be an arithmetic progression with a certain tolerance, such as: 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%.

[0044] Furthermore, in order to be able to compare with the treated cell test group, some indicator data of the target cells in the untreated cell test group under natural conditions can also be recorded, for example, the G1 phase proportion of the target cells under natural conditions, the Rb protein expression level, etc.

[0045] Alternatively, a cell control group can be prepared to be used as a reference for subsequent experiments.

[0046] S102, performing radiation operation on the multiple cell test groups, and determining cell biological indicators in each cell test group after the radiation operation.

[0047] After adjusting the G1 phase proportion of each cell test group, the cell test group is monitored for a certain period of time (such as 24-48 hours) to verify the stability of its G1 phase proportion. If large fluctuations are observed, the various parameters in the cell arrest treatment process can be further adjusted for optimization.

[0048] After determining that the G1 phase proportion of each cell test group is stable, each cell test group can be irradiated separately. After the irradiation operation, the cell biological indicators in each cell test group are recorded.

[0049] Among them, the mode and dosage of radiation operation can be set according to actual needs. For example, you can choose the radiation source (such as X-rays, gamma rays, heavy ions, etc.), radiation dose (such as 1Gy, 10Gy, etc.), and radiation method (such as single exposure, fractionated exposure, and other different radiation modes).

[0050] The recorded cell biological indicators may include: cell survival rate, cell cycle distribution, cell apoptosis rate, protein expression level, etc.

[0051] S103. Based on the G1 phase proportions and corresponding cell biological indicators of each cell test group, a data model is constructed to indicate the corresponding relationship between the G1 phase proportions of the target cells and the cell biological indicators, so as to evaluate the radiation sensitivity of the target cells.

[0052] According to the G1 phase proportion of each cell test group and the cell biological indicators recorded after radiation, a data model is constructed to reveal the corresponding relationship between the G1 phase proportion and the cell biological indicators. Through this model, the radiation sensitivity of the target cells can be further evaluated, and the effect of the G1 phase proportion on the radiation sensitivity of the target cells can be analyzed.

[0053] The constructed data model can be in various forms. For example, in one embodiment, a database records the correspondence between the G1 phase proportion and cell biological indicators; a neural network model or a pre-trained big data model is trained based on the measured G1 phase proportion and cell biological indicators as training samples, etc.

[0054] In the above embodiment, by performing cell cycle arrest treatment on the target cells, multiple cell test groups with different G1 phase proportions can be obtained, and then each cell test group is subjected to radiation operation respectively, and the cell biological indicators after the radiation operation are recorded, so that the relationship between the cell cycle and the cell radiation sensitivity can be quantitatively analyzed, and by constructing the corresponding data model, a reliable and accurate theoretical basis and data support are provided for subsequent research in the fields of cell biology, radiation biology, and personalized treatment.

[0055] In the above step S102, when recording the cell biological indicators of each cell test group after the radiation operation, it may include but is not limited to cell survival rate, cell proliferation rate, cell apoptosis rate, DNA damage degree, etc. In particular, cell survival rate is one of the important indicators reflecting the cell's response to radiation, which can directly reflect the cell's radiation sensitivity and tolerance.

[0056] Furthermore, based on the G1 phase proportion of each cell test group and the corresponding cell survival rate after radiation operation, a data model can be constructed to generate a quantitative correspondence between the G1 phase proportion of the target cells and the cell survival rate. This relationship can be formed into an accurate curve or function through fitting methods (such as regression analysis, support vector machine, etc.). For example, there may be a nonlinear relationship between the G1 phase proportion and the cell survival rate, and the data model can reveal the radiation response characteristics of cells under different G1 phase proportions.

[0057] By generating a correspondence between the G1 phase proportion of target cells and cell survival rate, the effect of G1 phase proportion on cell radiation sensitivity can be deeply analyzed. Specifically, researchers can clarify the contribution of G1 phase to radiation sensitivity by comparing the changes in cell survival rate under different G1 phase proportions, and then predict the radiation tolerance of cells at different cycle stages. This analysis not only helps to reveal the role of cell cycle regulatory mechanisms in radiation response, but also provides a quantifiable basis for the prediction of radiation sensitivity. By quantitatively analyzing the relationship between G1 phase proportion and cell survival rate and constructing a corresponding data model, it can not only reveal the intrinsic connection between cell cycle and radiation sensitivity, but also provide important support for the prediction of cell radiation sensitivity and the optimization of radiation therapy plans.

[0058] In one embodiment, when adjusting the G1 phase proportion of each cell test, doxycycline (Dox) can be added to multiple cell test groups of target cells to induce the expression level of retinoblastoma tumor suppressor protein (Rb) protein in the target cells under Dox treatment. Introducing the Dox-induced Rb allele into the target cell Rb knockout (KO) cell line can ensure that Dox-induced Rb protein expression is independent of cell size.

[0059] By adjusting the Dox concentration and / or treatment time in each cell test group, the Rb protein expression level and G1 phase proportion in the cell test group can be precisely controlled.

[0060] Different degrees of Rb protein expression can be induced by setting a Dox concentration gradient (such as 10, 20, 100, 500 ng / ml). It is expected that the G1 phase will be prolonged by Dox treatment, and different Dox concentrations may extend the G1 phase from slightly to about 30 hours.

[0061] The Dox treatment time for each cell test group can be adjusted within a certain time range, such as between 24 and 48 hours.

[0062] In the process of performing Dox treatment on each cell test group based on the preset Dox concentration and treatment time, the Rb protein expression level and G1 phase proportion in each cell test group can be monitored, and the Dox concentration and treatment time can be adjusted according to the monitoring results. For example, if the target G1 phase proportion in any cell test group is 40%-50%, but the current monitoring result shows that it is less than 40%, the Dox concentration can be appropriately increased or the treatment time can be extended.

[0063] There are many ways to monitor the expression level of Rb protein and the proportion of G1 phase, for example, Western Blotting, immunofluorescence staining, flow cytometry, enzyme-linked immunosorbent assay (ELISA), real-time cell monitoring system, etc.

[0064] The monitoring frequency can be set according to actual needs, for example, once every 4 hours.

[0065] Furthermore, based on the Dox concentration, treatment time, corresponding Rb protein expression level and G1 phase proportion in each cell test group, a relationship model between Dox concentration, treatment time, Rb protein expression level and G1 phase proportion can be constructed. For example, a corresponding relationship between Dox concentration and Rb protein expression level can be established; or a corresponding relationship between Rb protein concentration and G1 phase proportion can be established. After incorporating these relationship models into the data model, the effect of Dox concentration on Rb protein expression level or the effect of Rb protein concentration on G1 phase proportion can be further analyzed based on the data model.

[0066] Furthermore, by executing the above-mentioned Dox treatment process in multiple cycles and recording the experimental data each time, basic data can be accumulated for subsequent research and the experimental conditions can be further optimized.

[0067] In the above embodiment, by adjusting the Dox concentration and treatment time, the Rb protein expression level and the G1 phase proportion in the target cells can be accurately adjusted to achieve effective intervention in the cell cycle. By monitoring the expression of Rb protein and the G1 phase proportion at different concentrations and treatment times, the experimental conditions can be flexibly adjusted to ensure the repeatability and accuracy of the experimental results. By constructing a quantitative relationship model between Dox concentration, treatment time, Rb protein expression and G1 phase proportion, a theoretical basis is provided for subsequent experimental design and data analysis, which helps to deeply understand the specific effects of Dox treatment on the cell cycle.

[0068] In one embodiment, when the radiation operation is performed on the multiple cell test groups, each cell test group can be exposed to different doses of radiation. Specifically, multiple radiation dose gradients can be set in each cell test group, and the response of the target cells to different radiation doses can be observed, and the cell biological indicators under different radiation doses, such as cell survival rate, can be recorded.

[0069] The method for determining cell viability can be set according to actual needs. For example, a clone formation experiment or a cell viability analysis (such as MTS assay) can be used to measure the cell viability after radiation operation.

[0070] Furthermore, a relationship model between radiation dose, G1 phase proportion, and cell survival rate can be established and incorporated into the data model, so that the data model can be used to determine the impact of the cell cycle on the radiation sensitivity of target cells under different radiation doses, especially the relationship between the cell cycle and cell survival rate.

[0071] By setting a radiation dose gradient in multiple cell test groups, the effects of different radiation doses on cell biological indicators (such as cell survival rate) can be systematically studied, providing data support for the evaluation of radiation sensitivity.

[0072] The target cells in the disclosed embodiments can be selected according to actual needs. For example, in one embodiment, tumor cells can be selected as target cells, and finally a data model of tumor cells can be constructed to explore the role of its cell cycle in tumor radiotherapy, which provides an important experimental basis for the targeted regulation of radiotherapy, lays a foundation for clinical personalized treatment, and improves the survival prognosis of patients. It can also provide theoretical support for the future development of cell cycle regulation drugs or new treatment strategies.

[0073] In one embodiment, a head and neck tumor cell line (such as UM-SCC-1 or UM-SCC-47) can be selected as a target cell, which is representative in head and neck cancer radiotherapy research.

[0074] Figure 21 is a complete flow chart of a method for determining cell radiation sensitivity provided by an exemplary embodiment, the steps of which include:

[0075] 1. Cell preparation and initial condition confirmation

[0076] 1.1 Select cell line

[0077] Choose a head and neck tumor cell line (such as UM-SCC-1 or UM-SCC-47) that is representative in head and neck cancer radiotherapy research.

[0078] 1.2 Cell culture

[0079] The cells were cultured under standard culture conditions (37° C., 5% CO 2 , 10% fetal bovine serum) to ensure that the cells were in the logarithmic growth phase to ensure the stability of the cell cycle.

[0080] 1.3 Establishment of control group

[0081] An untreated control group was established to record the proportion of G1 phase cells in the natural state, and to measure the basal expression level of Rb protein as a reference for subsequent experiments.

[0082] 2. Rb protein induction and Dox concentration control

[0083] 2.1 Dox-induced Rb protein expression level

[0084] Introduction of a Dox-inducible Rb allele into an RB knockout (KO) head and neck tumor cell line ensured that induction of Rb protein was independent of cell size.

[0085] 2.2 Dox concentration gradient

[0086] A Dox concentration gradient (10, 20, 100, 500 ng / ml) was set, and it was expected to prolong the G1 phase (from a slight extension to about 30 hours).

[0087] 2.3 Processing time

[0088] Under treatment with different concentrations of Dox (24-48 hours), the changes in the expression level of Rb protein were observed.

[0089] 2.4 Real-time monitoring

[0090] 2.4.1 Flow cytometry and Rb antibody labeling were used to monitor the Rb protein expression level and the proportion of G1 phase cells in real time every 4 hours, and the effects of different Dox concentrations on cell status were dynamically recorded.

[0091] 2.4.2 Adjust Dox dosage and treatment time in real time according to monitoring data.

[0092] 2.4.3 Short-term feedback loop: Through at least two rounds of feedback (monitoring-adjustment-verification), the Dox concentration gradient of each experimental group is optimized to ensure that the expected Rb expression level and G1 phase proportion are achieved, and to accumulate data for subsequent experiments.

[0093] 2.5 Target G1 phase proportion

[0094] 2.5.1 Set the target G1 phase percentage to 20%-95%, with each 5% as an interval, ensuring that the tolerance is within ±5%. Optimize the G1 phase cell percentage by dynamically adjusting the Dox concentration and treatment time.

[0095] 2.5.2 Determine the optimal treatment time range under different Dox concentrations to ensure that the proportion of G1 phase cells remains stable within this time range to avoid cell decline or cycle disorder.

[0096] 3. G1 phase proportion measurement and data collection

[0097] 3.1 Cell cycle analysis

[0098] Flow cytometry or BrdU / PI labeling was used to analyze the cell cycle after treatment with different Dox concentrations, and the proportion of cells in the G1 phase was recorded.

[0099] 3.2 Determination of Rb protein expression level

[0100] The expression level of Rb protein at different Dox concentrations was quantitatively analyzed by Western blot or immunofluorescence to verify the Dox-induced effect.

[0101] 3.3 Generate Relational Model

[0102] The relationship between Rb protein concentration and the proportion of cells in the G1 phase was recorded, and the concentration-proportion curve was drawn to analyze the effect of Rb protein concentration on the proportion of cells in the G1 phase.

[0103] 3.4 Verification of Steady-State of G1 Proportion

[0104] 3.4.1 Under the optimized Dox treatment conditions, continuously monitor (24-48 hours) the stability of the G1 phase proportion of each experimental group. If large fluctuations are found, optimization can be performed by adjusting the treatment time or Dox concentration.

[0105] 3.4.2 The steady-state G1 phase proportion, Rb protein concentration and cell morphology changes under each group of conditions were recorded, and a standardized curve of Dox concentration, treatment time and G1 phase proportion was constructed.

[0106] 3.5 Experimental cycle optimization

[0107] 3.5.1 According to different target G1 phase proportions, repeatedly optimize the experimental parameters to ensure that a repeatable concentration gradient and treatment time range are obtained, and the G1 phase proportion interval is controlled within 2% as much as possible.

[0108] 3.5.2 Integrate the results of multiple rounds of experiments, analyze the dynamic effect of Dox concentration on the proportion of G1 phase, and further optimize the concentration gradient standardization scheme.

[0109] 4. Radiation operation and data analysis

[0110] 4.1 Radiation Operation

[0111] Radiation treatment (common dose range: 2-8 Gy) was performed on each Dox-treated group, and multiple dose gradients were set to observe the cell response at different doses.

[0112] 4.2 Cell viability determination

[0113] The cell survival rate after irradiation was evaluated by clone formation assay or MTS assay, and the survival rate curve was drawn.

[0114] 4.3 Relationship between G1 phase proportion and cell survival rate

[0115] The different G1 phase proportions were compared with the radiation survival rate, and the relationship curve between the G1 phase proportion and the survival rate was drawn.

[0116] 5. Data model establishment

[0117] 5.1 Experimental data collection

[0118] After the radiation experiment, the survival rate data of each treatment group were collected and compared with the proportion of G1 phase to explore the relationship between the two.

[0119] 5.2 Establishing the Relationship Model

[0120] Generate a curve showing the relationship between survival rate and G1 phase proportion, and generate a data model.

[0121] 5.3 Cell cycle state inference

[0122] Based on the relationship between survival rate and G1 phase percentage, the cell cycle status can be inferred. For example, low survival rate and high G1 phase percentage may indicate that cells stay in G1 phase too long, reducing radiation tolerance.

[0123] 5.4 Result verification and adjustment

[0124] According to the inference results, the experimental design can be optimized. If the cell survival rate is low under a specific G1 phase ratio, the G1 phase cell ratio can be optimized by adjusting the Dox concentration or treatment time to improve the radiotherapy effect.

[0125] 6. Subsequent application of data model

[0126] 6.1 Cancer treatment

[0127] The relationship curve between the proportion of G1 phase and survival rate provides an important basis for cancer treatment. Researchers can adjust the radiotherapy dose and chemotherapy drug use according to the proportion of G1 phase cells to achieve personalized treatment plans, optimize efficacy and reduce normal cell damage.

[0128] 6.2 Stem Cell Research

[0129] In cancer stem cell research, the regulation of the proportion of G1 phase cells helps analyze stem cell proliferation and drug resistance. By monitoring changes in the proportion of G1 phase cells, accurate data can be provided for targeted treatment of cancer stem cells.

[0130] 6.3 Drug Screening and Evaluation

[0131] The relationship curve between the G1 phase proportion and the survival rate provides a quantitative indicator for drug screening, helps evaluate the anti-proliferative effect of drugs in the G1 phase, screens out potential anti-cancer drugs, and provides a basis for the evaluation of the efficacy and side effects of new drugs.

[0132] Please refer to Figure 3 , Figure 3 A cell radiation sensitivity determination device is shown to implement the technical solution of this specification. The cell radiation sensitivity determination device may include: a cell cycle adjustment module 301 , a radiation module 302 and a data analysis module 303 .

[0133] The cell cycle adjustment module 301 is used to use a preset cell cycle arrest method to process multiple cell test groups of target cells respectively, and obtain multiple cell test groups with different G1 phase proportions; wherein the G1 phase proportion is used to indicate the proportion of cells in the G1 phase in the cell test group; the radiation module 302 is used to perform radiation operations on the multiple cell test groups, and determine the cell biological indicators in each cell test group after the radiation operation; the data analysis module 303 is used to construct a data model for indicating the correspondence between the G1 phase proportion of the target cells and the cell biological indicators based on the G1 phase proportion of each cell test group and the corresponding cell biological indicators, so as to evaluate the radiation sensitivity of the target cells.

[0134] Furthermore, the cell biological index includes cell survival rate.

[0135] Furthermore, the data analysis module 303 is used to construct a relationship curve between the G1 phase proportion of the target cells and the cell survival rate.

[0136] Furthermore, the cell cycle adjustment module 301 is used to add doxycycline Dox to multiple cell test groups of target cells to induce the expression level of retinoblastoma tumor suppressor protein Rb protein in the target cells under Dox treatment; by adjusting the Dox concentration and / or treatment time in each cell test group, the G1 phase proportion is adjusted with different Rb protein expression levels.

[0137] Furthermore, the data analysis module 303 is also used to construct a corresponding relationship between the Dox concentration and the Rb protein expression level based on the Dox concentration and the corresponding Rb protein expression level in each cell test group.

[0138] Furthermore, the data analysis module 303 is also used to construct a corresponding relationship between the Rb protein concentration and the G1 phase proportion of the target cells based on the Rb protein concentration and the corresponding G1 phase proportion of each cell test group.

[0139] Furthermore, the radiation module 302 is used to expose each cell test group to different doses of radiation.

[0140] Furthermore, the data model is also used to indicate the corresponding relationship between the cell cycle and the cell survival rate of the target cells under different radiation doses.

[0141] Furthermore, the target cells are tumor cells.

[0142] Furthermore, the tumor cells are head and neck tumor cell lines.

[0143] Based on the same concept as the above method, this specification also provides an electronic device, including: a processor; a memory for storing processor executable instructions; wherein the processor implements the steps of the method described in any of the above embodiments by running the executable instructions.

[0144] Based on the same concept as the above method, this specification also provides a computer-readable storage medium on which computer instructions are stored. When the instructions are executed by a processor, the steps of the method described in any of the above embodiments are implemented.

[0145] Based on the same concept as the above method, this specification also provides a computer program product, including a computer program / instruction, which implements the steps of the method described in any of the above embodiments when executed by a processor.

Claims

1. A method for determining cell radiation sensitivity, characterized in that: The method comprises: A preset cell cycle arrest method is used to process multiple cell test groups of target cells respectively, so as to obtain multiple cell test groups with different G1 phase proportions; wherein the G1 phase proportion is used to indicate the proportion of cells in the G1 phase in the cell test group; Performing radiation operation on the multiple cell test groups, and determining cell biological indicators in each cell test group after the radiation operation; Based on the G1 phase proportions and corresponding cell biological indicators of each cell test group, a data model for indicating the corresponding relationship between the G1 phase proportions of the target cells and the cell biological indicators is constructed to evaluate the radiation sensitivity of the target cells.

2. The method according to claim 1, characterized in that The cell biological index includes cell survival rate.

3. The method according to claim 2, characterized in that The corresponding relationship between the G1 phase proportion of the target cells and the cell biological indexes includes: a relationship curve between the G1 phase proportion of the target cells and the cell survival rate.

4. The method according to claim 1, characterized in that: The method of using a preset cell cycle arrest method to process multiple cell test groups of target cells respectively includes: Adding doxycycline Dox to multiple cell test groups of target cells to induce the expression level of retinoblastoma tumor suppressor protein Rb protein in the target cells under Dox treatment; By adjusting the Dox concentration and / or treatment time in each cell test group, the G1 phase proportion was adjusted with different Rb protein expression levels.

5. The method according to claim 4, characterized in that The method further comprises: Based on the Dox concentration and the corresponding Rb protein expression level in each cell test group, the corresponding relationship between the Dox concentration and the Rb protein expression level was constructed.

6. The method according to claim 5, characterized in that The method further comprises: Based on the Rb protein concentration and the corresponding G1 phase proportion of each cell test group, the corresponding relationship between the Rb protein concentration and the G1 phase proportion of the target cell is constructed.

7. The method according to claim 1, characterized in that The irradiation operation on the plurality of cell test groups comprises: Each test group of cells was exposed to a different dose of radiation.

8. The method according to claim 1, characterized in that: The data model is also used to indicate the corresponding relationship between the cell cycle and the cell survival rate of the target cells under different radiation doses.

9. The method according to claim 1, characterized in that: The target cells are tumor cells.

10. The method according to claim 9, characterized in that The tumor cells are head and neck tumor cell lines.

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