The Correlation Between Tudor-SN Protein And Radiation Resistance Of Cervical Cancer

Objectives: Cervical cancer is one of the most common gynecological malignancies in the world,the cure rate of postoperative women in middle and late stage women is very low.At present,radiotherapy is one of the main methods of medical treatment for cervical cancer.The cure effect of cervical cancer has this obvious effect,but after radiotherapy,cervical cancer still has the problems of high recurrence,high metastasis and radiation resistance.These problems are the main obstacle of clinical radiotherapy cancer.Therefore,it is of great significance to explore the complex biological activity of radiation resistance of tumor cells for radiotherapy of cancer.The sensitivity of tumor radiation depends on the repair ability of DNA damage,while the tumor cells with radiation resistance have enhanced DNA damage repair ability.The radiation resistance of tumor is mainly manifested in three aspects: DNA damage repair response,cell cycle arrest and inhibition of cell apoptosis.In recent years,it has been reported that Tudor-SN protein can be modified by ADP ribosomal polymerase family proteins.In addition,we found that the ADP ribosylation modification was related to the repair of DNA damage and energy metabolism.In addition,the lab early found that Tudor-SN was involved in the regulation of cell cycle,and mass spectrometric detection experiment detected Tudor-SN protein and PARP-1 repair protein binding and affect the sensitivity of tumor cells to radiation.Therefore,this topic is mainly based on the original research to explore the Tudor-SN protein and cervical cancer radiation resistance correlation,and draws the differential expression profile of cervical cancer radiation resistance strain by proteomics experiment.Combined with bioinformatics to analyze the potential role of Tudor-SN protein in radiation resistance of cervical cancer.Methods:The research is divided into two parts: The first part is to construct a stable He La-R cell line with radiation resistance.This part is validated by the following four aspects: 1)Analyze the radiosensitivity of cancer cells by clonogenic assay;2)Detect the proliferation ability of cancer cells by CCK-8;3)Detect apoptosis by flow cytometry and Western blotting were used to detect the apoptosis markers.4)The distribution of cell cycle was detected by flow cytometry and the cell cycle markers were detected by Western blotting.The second part of the study is the correlation between Tudor-SN protein and radiation resistance of cervical cancer.On the basis of He La-R cells,we constructed Tudor-SN protein knockout cell stable cell line He La-R-KO.We performed proteomic analysis of He La cells,He La-R cells and He La-R-KO cells,identified differentially expressed proteins between cells,systematic biological analysis of protein containing quantitative information.Results: Part1,The intermittent irradiation induction method was used to construct a stable cervical cancer cell strain He La-R with radiation resistance.Compared with the parental He La cells,He La-R cells have lower sensitivity and faster proliferation after irradiation.The apoptosis rate of He La-R cells was significantly lower than that of He La cells by flow cytometry and the expression of apoptosis markers was also inhibited.In addition,cell cycle experiments showed that He La-R cells had significantly higher G2/M block distribution,the expression of cycle-related markers increased.Part2,Knockout of Tudor-SN protein increased the apoptosis rate and radiosensitivity of He La-R cells.Through proteomic analysis of He La cells,He La-R cells and He La-R-KO cells,a total of 6469 proteins were identified in this study,of which 5903 proteins contained quantitative information.When t-test p-value <0.05 was used as the standard,1.3 times the change threshold value,574 proteins in He La-R cells/He La cells were up-regulated and 856 proteins were down-regulated in the quantified protein.In He La-R-KO/He La-R group,104 proteins were up-regulated and 980 proteins were down-regulated.The preliminary bioinformatics analysis of differentially expressed proteins laid a foundation for the study of subsequent radiation resistance.Conclusions: 1.The intermittent irradiation induction method was used to construct a stable cervical cancer cell strain He La-R with radiation resistance.2.He La-R cells have stronger radiation resistance and cell proliferation ability than He La cells,and have obvious G2/M block and anti-apoptotic ability after radiotherapy.3.The proteomic data of He La-R cells and He La-R-KO cells of He La cells were systematically analyzed by bioinformatics,including protein annotation,functional classification,functional enrichment and cluster analysis based on functional enrichment.for further research on Tudor-SN protein intervention in cervical cancer radiation resistance provides reference direction.