Method for determining cell radiosensitivity

Abstract

The one or more embodiments of the specification provide a cell radiosensitivity determination method, comprising: adopting a preset cell cycle arrest method to process a plurality of cell test groups of target cells respectively, 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 a radiation operation on the plurality of cell test groups, and determining a cell biological index in each cell test group after the radiation operation; based on 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 and the cell biological index of the target cell, for evaluating the radiation sensitivity of the target cell.

Description

Technical Field

[0001] This specification relates to one or more embodiments in the field of biotechnology, and more particularly to a method for determining cell radiation sensitivity. Background Technology

[0002] Cellular radiation response is typically closely related to its stage in the cell cycle, and different cell cycle stages exhibit significant differences in radiation sensitivity. Therefore, exploring the relationship between the cell cycle and the survival rate of tumor cells after radiation exposure is of significant research value.

[0003] Related research mainly employs methods such as clonogenic assays, heavy ion irradiation based on cell cycle synchronization, linear quadratic (LQ) models, and flow cytometry. However, these techniques each have their own limitations, such as long experimental cycles, complex operation, insufficient sensitivity, or inability to comprehensively consider factors such as the cellular microenvironment. Therefore, in-depth quantitative research on the relationship between cell cycle and cellular radiation sensitivity remains lacking. Summary of the Invention

[0004] In view of the above, 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] Multiple cell test groups of target cells were processed using a preset cell cycle arrest method to obtain multiple cell test groups with different proportions of G1 phase; wherein, the proportion of G1 phase is used to indicate the proportion of cells in the cell test group that are in the G1 phase.

[0007] Irradiation was performed on the multiple cell test groups, and cell biological indicators in each cell test group were determined after the irradiation.

[0008] Based on the proportion of G1 phase and corresponding cell biological indicators of each cell test group, a data model is constructed to indicate the correspondence between the proportion of G1 phase and cell biological indicators of the target cells, which is used to evaluate the radiation sensitivity of the target cells.

[0009] Furthermore, the cell biology indicators include cell viability.

[0010] Furthermore, the correspondence between the proportion of target cells in G1 phase and cell biological indicators includes: the relationship curve between the proportion of target cells in G1 phase and cell viability.

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

[0012] Doxycycline (Dox) was added to multiple cell test groups of target cells to induce the expression level of retinoblastoma tumor suppressor protein Rb in target cells under Dox treatment;

[0013] The proportion of G1 phase was adjusted by varying the Dox concentration and / or treatment time in each cell test group to achieve different Rb protein expression levels.

[0014] Furthermore, the method also includes:

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

[0016] Furthermore, the method also includes:

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

[0018] Furthermore, the irradiation process performed on the plurality of cell test groups includes:

[0019] Each cell test group was exposed to different doses of radiation.

[0020] Furthermore, the data model is also used to indicate the correspondence between the cell cycle and 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] As can be seen from the above embodiments, this specification can obtain multiple cell test groups with different G1 phase proportions by performing cell cycle arrest on target cells. Then, each cell test group is subjected to radiation operation, and the cell biological indicators after radiation operation are recorded. This allows for quantitative analysis of the relationship between cell cycle and cell radiation sensitivity. Furthermore, by constructing corresponding data models, it provides a reliable and accurate theoretical basis and data support for subsequent research in cell biology, radiation biology, personalized therapy, and other fields. Attached Figure Description

[0024] Figure 1 This is a flowchart of a method for determining cell radiation sensitivity, provided in an exemplary embodiment.

[0025] Figure 2 This is a schematic flowchart of a method for determining cell radiation sensitivity, provided in an exemplary embodiment.

[0026] Figure 3 This is a block diagram of a cell radiation sensitivity determination device provided in an exemplary embodiment. Detailed Implementation

[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, data stored, data displayed, etc.) involved in this manual are all information and data authorized by the user or fully authorized by all parties. Furthermore, the collection, use and processing of related data must comply with the relevant laws, regulations and standards of the relevant countries and regions, and corresponding operation portals are provided for users to choose to authorize or refuse.

[0028] Cellular radiation response is typically closely related to its stage in the cell cycle, and different cell cycle stages exhibit significant differences in radiation sensitivity. Therefore, exploring the relationship between the cell cycle and the survival rate of tumor cells after radiation exposure is of significant research value.

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

[0030] Clonogenesis assays: These assays obtain radiation biological parameters by directly measuring cell viability after irradiation and plotting survival curves. However, this method suffers from a long experimental cycle, typically requiring days to weeks to observe cell viability, leading to data acquisition delays. Furthermore, the sensitivity of measurements is low at low doses of radiation.

[0031] Heavy ion irradiation based on cell cycle synchronization: This method models cell dynamics using experimental data to track dynamic changes in cells under different irradiation protocols. The drawbacks of this method are that stress responses may occur during cell cycle synchronization, altering cell radiation sensitivity, and the synchronization effect is difficult to guarantee consistently between different experiments, affecting data reproducibility and reliability.

[0032] Linear Quadratic (LQ) models quantify cellular radiation sensitivity using parameters α and β and fit cell survival curves. However, LQ models assume linear and quadratic effects in radiation responses, which may prevent them from accurately reflecting actual biological responses in complex situations such as high doses or fractionated doses. Furthermore, this model requires extensive data and cannot account for individual variability.

[0033] Flow cytometry: Used to accurately determine cell cycle distribution, especially labeled S-phase cells, to analyze the cell population's response to radiation at different phases. Although flow cytometry can provide this data, it is complex to operate, requires multiple labeling for result interpretation, and primarily analyzes the transient state of cells, unable to provide long-term survival information.

[0034] In summary, each of the above techniques has its limitations, such as long experimental cycles, complex operation, insufficient sensitivity, or difficulty in comprehensively considering factors such as the cellular microenvironment. These shortcomings affect the accurate assessment of cellular radiation sensitivity.

[0035] The cell cycle can be divided into two main parts: interphase and mitosis (M). Interphase is further divided into three subphases: pre-interphase (Gap 1, G1), DNA synthesis (S), and post-interphase (Gap 2, G2). When exposed to radiation, G1 phase cells are generally more tolerant, S phase cells have relatively lower radiation sensitivity, while G2 and M phase cells are most sensitive to radiation.

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

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

[0038] Please see Figure 1 , Figure 1 This is an exemplary embodiment of a method for determining cellular radiation sensitivity. The method may include the following steps.

[0039] S101. Multiple cell test groups of target cells are processed using a preset cell cycle arrest method 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 implementation, multiple cell test sets can be pre-prepared. For example, target cells are cultured under preset culture conditions (e.g., 37°C, 5% CO2, 10% fetal bovine serum) to obtain multiple cell test sets. To ensure the stability and reproducibility of experimental results, the pre-prepared cell test sets should ensure that the target cells are in the logarithmic growth phase. During this phase, the cell division rate and proliferation rate are relatively constant, the distribution of each cell cycle stage is balanced, and it is not easily affected by fluctuations in the external environment. Therefore, ensuring that 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 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 was treated using a pre-defined cell cycle arrest method to adjust the proportion of target cells in the G1 phase in each cell test group, so as to obtain a series of cell test groups with different proportions of G1 phase.

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

[0043] After adjustment, the percentages of G1 phase in multiple cell test groups can be randomly distributed or precisely controlled within predetermined intervals, such as 30%-40%, 40%-50%, etc. They can also be further controlled to multiple specified target G1 phase percentages, such as 20%, 30%, 40%, etc. These target G1 phase percentages can be an arithmetic sequence with a certain common difference, for example: 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%.

[0044] Furthermore, in order 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, such as the proportion of target cells in G1 phase and the expression level of Rb protein under natural conditions.

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

[0046] S102. Perform irradiation on the plurality of cell test groups, and determine the cell biological indicators in each cell test group after the irradiation.

[0047] After adjusting the proportion of G1 phase in each cell test group, steady-state monitoring was performed on the cell test groups for a certain period of time (e.g., 24-48 hours) to verify the stability of their G1 phase proportion. If large fluctuations are observed, further adjustments can be made to the various parameters in the cell arrest treatment process for optimization.

[0048] After confirming that the proportion of cells in the G1 phase is stable in each cell test group, irradiation can be performed on each cell test group separately. After irradiation, cell biological indicators in each cell test group are recorded.

[0049] The method and dose of radiation operation can be set according to actual needs. For example, the radiation source (such as X-ray, gamma ray, heavy ion, etc.), radiation dose (such as 1 Gy, 10 Gy, etc.), and radiation method (such as single exposure, multiple exposure, etc.) can be selected.

[0050] Recorded cell biology indicators may include: cell viability, cell cycle distribution, apoptosis rate, protein expression level, etc.

[0051] S103. Based on the proportion of G1 phase and corresponding cell biological indicators of each cell test group, construct a data model to indicate the correspondence between the proportion of G1 phase and cell biological indicators of the target cells, and use it to evaluate the radiation sensitivity of the target cells.

[0052] Based on the proportion of G1 phase cells in each cell test group and the cell biological indicators recorded after irradiation, a data model was constructed to reveal the correlation between the proportion of G1 phase cells and cell biological indicators. This model can be used to further assess the radiation sensitivity of target cells and analyze the impact of the proportion of G1 phase cells on their radiation sensitivity.

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

[0054] In the above embodiments, by performing cell cycle arrest on the target cells, multiple cell test groups with different G1 phase proportions can be obtained. Then, each cell test group is subjected to radiation operation, and the cell biological indicators after radiation operation are recorded. This allows for quantitative analysis of the relationship between cell cycle and cell radiation sensitivity. Furthermore, by constructing corresponding data models, reliable and accurate theoretical basis and data support are provided for subsequent research in cell biology, radiation biology, personalized therapy, and other fields.

[0055] In step S102 above, when recording cell biological indicators for each cell test group after radiation treatment, these may include, but are not limited to, cell viability, cell proliferation rate, apoptosis rate, and degree of DNA damage. Cell viability, in particular, is one of the important indicators reflecting the cell's response to radiation, directly reflecting the cell's radiation sensitivity and tolerance.

[0056] Furthermore, based on the proportion of cells in G1 phase in each cell test group and the corresponding cell viability after irradiation, a data model can be constructed to generate a quantitative correlation between the proportion of target cells in G1 phase and cell viability. This relationship can be fitted using methods such as regression analysis or support vector machines to form an accurate curve or function. For example, the relationship between the proportion of cells in G1 phase and cell viability may exhibit a non-linear relationship, and the data model can reveal the radiation response characteristics of cells under different proportions of cells in G1 phase.

[0057] By analyzing the correlation between the proportion of target cells in G1 phase and cell viability, the impact of G1 phase proportion on cellular radiation sensitivity can be further investigated. Specifically, researchers can compare changes in cell viability under different G1 phase proportions to clarify the contribution of G1 phase to radiation sensitivity, thereby predicting the radiation tolerance of cells at different stages of the cell cycle. This analysis not only helps to reveal the role of cell cycle regulation mechanisms in radiation response but also provides a quantifiable basis for predicting radiation sensitivity. Quantitative analysis of the relationship between G1 phase proportion and cell viability, and the construction of corresponding data models, can not only reveal the intrinsic link between the cell cycle and radiation sensitivity but also provide important support for predicting cellular radiation sensitivity and optimizing radiation therapy regimens.

[0058] In one implementation, when adjusting the proportion of G1 phase in each cell assay, doxycycline (Dox) can be added to multiple cell assay groups of the target cells to induce the expression level of retinoblastoma protein (Rb) in the target cells under Dox treatment. Introducing the Dox-induced Rb allele into the Rb knockout (KO) cell line of the target cells ensures 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 expression level of Rb protein and the proportion of G1 phase in the cell test group can be precisely controlled.

[0060] Different levels of Rb protein expression can be induced by setting Dox concentration gradients (e.g., 10, 20, 100, 500 ng / ml). It is expected that the G1 phase will be prolonged by Dox treatment, and different Dox concentrations may prolong 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] During the Dox treatment of each cell test group based on the preset Dox concentration and treatment time, the expression level of Rb protein and the proportion of G1 phase in each cell test group can be monitored. Based on the monitoring results, the Dox concentration and treatment time can be adjusted. For example, if the target G1 phase proportion in any cell test group is 40%-50%, but the current monitoring results show that it is lower than 40%, the Dox concentration can be appropriately increased or the treatment time can be extended.

[0063] There are various methods for monitoring Rb protein expression levels and the proportion of G1 phase, such as Western blotting, immunofluorescence staining, flow cytometry, enzyme-linked immunosorbent assay (ELISA), and real-time cell monitoring systems.

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

[0065] Furthermore, based on the Dox concentration, treatment time, corresponding Rb protein expression level, and G1 phase percentage in each cell test group, a relationship model can be constructed between Dox concentration, treatment time, Rb protein expression level, and G1 phase percentage. For example, a correspondence can be established between Dox concentration and Rb protein expression level; or a correspondence can be established between Rb protein concentration and G1 phase percentage. 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 percentage, can be further analyzed based on this data model.

[0066] Furthermore, by repeatedly executing the above Dox processing procedure and recording the experimental data each time, basic data can be accumulated for subsequent research, and experimental conditions can be further optimized.

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

[0068] In one implementation, when performing radiation operations on the plurality of 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, recording cell biological indicators such as cell viability under different radiation doses.

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

[0070] Furthermore, a relationship model can be established between radiation dose, G1 phase percentage, and cell viability, and incorporated into the data model. This data model can then be used to determine the impact of cell cycle on the radiation sensitivity of target cells under different radiation doses, particularly the relationship between cell cycle and cell viability.

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

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

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

[0074] Figure 2This is a schematic diagram of a complete process for determining cell radiation sensitivity, provided in an exemplary embodiment, the steps of which include:

[0075] 1. Cell preparation and initial condition confirmation

[0076] 1.1 Selection of cell lines

[0077] Select head and neck tumor cell lines (such as UM-SCC-1 or UM-SCC-47), which are representative of head and neck cancer radiotherapy research.

[0078] 1.2 Cell Culture

[0079] Cells were cultured under standard culture conditions (37°C, 5% CO2, 10% fetal bovine serum) to ensure that the cells were in the logarithmic growth phase, thus guaranteeing cell cycle stability.

[0080] 1.3 Establishment of Control Group

[0081] An untreated control group was set up to record the percentage of cells in the G1 phase under natural conditions 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] Introducing Dox-induced Rb alleles into RB knockout (KO) head and neck tumor cell lines ensures that Rb protein induction is independent of cell size.

[0085] 2.2 Dox concentration gradient

[0086] Set up Dox concentration gradients (10, 20, 100, 500 ng / ml) to prolong the G1 phase (from slightly prolonged to approximately 30 hours).

[0087] 2.3 Processing time

[0088] Changes in Rb protein expression levels were observed under different concentrations of Dox treatment (24-48 hours).

[0089] 2.4 Real-time monitoring

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

[0091] 2.4.2 Adjust the Dox dosage and treatment time in real time based on monitoring data.

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

[0093] 2.5 Target G1 phase proportion

[0094] 2.5.1 The target G1 phase percentage was set at 20%-95%, with each 5% interval ensuring a tolerance within ±5%. The G1 phase cell percentage was optimized by dynamically adjusting the Dox concentration and treatment time.

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

[0096] 3. Measurement and Data Collection of the Proportion of G1 Period

[0097] 3.1 Cell cycle analysis

[0098] The cell cycle was analyzed using flow cytometry or BrdU / PI labeling, and the proportion of cells in 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 Generative Relationship Model

[0102] Record the relationship between Rb protein concentration and the proportion of cells in G1 phase, plot the concentration-proportion curve, and analyze the effect of Rb protein concentration on the proportion of cells in G1 phase.

[0103] 3.4G1 phase steady-state verification

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

[0105] 3.4.2 Record the percentage of steady-state G1 phase, Rb protein concentration and cell morphology changes under each group of conditions, and construct a standardized curve of Dox concentration, treatment time and percentage of G1 phase.

[0106] 3.5 Experimental Cycle Optimization

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

[0108] 3.5.2 Integrate the results of multiple rounds of experiments, analyze the dynamic impact 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 was performed on each Dox treatment group (common dose range: 2-8 Gy), and multiple dose gradients were set to observe cell responses at different doses.

[0112] 4.2 Cell viability assay

[0113] Cell viability after irradiation was assessed using clonogenic assays or MTS assays, and viability curves were plotted.

[0114] 4.3 Relationship between the proportion of cells in G1 phase and cell viability

[0115] By comparing the proportion of different G1 phases with the radiation survival rate, a curve showing the relationship between the proportion of G1 phases and the survival rate was plotted.

[0116] 5. Data Model Establishment

[0117] 5.1 Experimental Data Collection

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

[0119] 5.2 Establishment of the Relational Model

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

[0121] 5.3 Cell cycle state inference

[0122] By analyzing the relationship between cell survival rate and the percentage of cells in G1 phase, we can infer the cell cycle state. For example, a low survival rate and a high percentage of cells in G1 phase may indicate that cells remain in G1 phase for too long, reducing their tolerance to radiation.

[0123] 5.4 Result Verification and Adjustment

[0124] Based on the back-calculation results, optimize the experimental design. If it is found that the cell survival rate is low at a specific proportion of G1 phase cells, the proportion of G1 phase cells can be optimized by adjusting the Dox concentration or treatment time to improve the radiotherapy effect.

[0125] 6. Subsequent applications of the data model

[0126] 6.1 Cancer Treatment

[0127] The relationship between the proportion of G1-phase cells and survival rate provides important data for cancer treatment. Researchers can adjust radiotherapy doses and chemotherapy drugs based on the proportion of G1-phase cells to achieve personalized treatment plans, optimize efficacy, and reduce damage to normal cells.

[0128] 6.2 Stem Cell Research

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

[0130] 6.3 Drug Screening and Evaluation

[0131] The curve showing the relationship between the proportion of G1-phase cells and survival rate provides a quantitative indicator for drug screening, helps to assess the antiproliferative effect of drugs in the G1 phase, screens out potential anticancer drugs, and provides a basis for evaluating 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 process multiple cell test groups of target cells using a preset cell cycle arrest method 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; 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 based on the G1 phase proportion and corresponding cell biological indicators of each cell test group to indicate the correspondence between the G1 phase proportion and cell biological indicators of the target cells, and to evaluate the radiation sensitivity of the target cells.

[0134] Furthermore, the cell biology indicators include cell viability.

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

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

[0137] Furthermore, the data analysis module 303 is also used to construct the correspondence between the Dox concentration and the corresponding 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 the correspondence between the Rb protein concentration and the G1 phase ratio of the target cells based on the Rb protein concentration and the corresponding G1 phase ratio 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 correspondence between the cell cycle and 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 methods described above, this specification also provides an electronic device, including: a processor; a memory for storing processor-executable instructions; wherein the processor performs the steps of the method as described in any of the above embodiments by executing the executable instructions.

[0144] Based on the same concept as the methods described above, this specification also provides a computer-readable storage medium having computer instructions stored thereon that, when executed by a processor, implement the steps of the methods as described in any of the above embodiments.

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

Claims

1. A method for determining cell radiation sensitivity, characterized in that, The method includes: Multiple cell test groups of target cells were treated using a pre-defined cell cycle arrest method 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; the treatment of multiple cell test groups of target cells using the pre-defined cell cycle arrest method includes: adding doxycycline (Dox) to the multiple cell test groups of target cells to induce the expression level of the retinoblastoma tumor suppressor protein Rb in the target cells under Dox treatment; adjusting the Dox concentration and / or treatment time in each cell test group to adjust the G1 phase proportion to different Rb protein expression levels; Irradiation was performed on the multiple cell test groups, and cell biological indicators in each cell test group were determined after the irradiation. Based on the proportion of G1 phase and corresponding cell biological indicators of each cell test group, a data model is constructed to indicate the correspondence between the proportion of G1 phase and cell biological indicators of the target cells, which is used to evaluate the radiation sensitivity of the target cells.

2. The method according to claim 1, characterized in that, The cell biology indicators include cell viability.

3. The method according to claim 2, characterized in that, The correspondence between the proportion of target cells in G1 phase and cell biological indicators includes: the relationship curve between the proportion of target cells in G1 phase and cell viability.

4. The method according to claim 1, characterized in that, The method further includes: Based on the Dox concentration and the corresponding Rb protein expression level in each cell test group, the correspondence between the Dox concentration and the Rb protein expression level was constructed.

5. The method according to claim 4, characterized in that, The method further includes: Based on the Rb protein concentration and the corresponding proportion of G1 phase in each cell test group, the correspondence between the Rb protein concentration and the proportion of G1 phase in the target cells was constructed.

6. The method according to claim 1, characterized in that, The radiation operation performed on the multiple cell test groups includes: Each cell test group was exposed to different doses of radiation.

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

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

9. The method according to claim 8, characterized in that, The tumor cells are head and neck tumor cell lines.

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