CDK13 Interacts With E2F5 To Regulate Prostate Cancer Research On Proliferation And Mechanism Of Action

Prostate cancer belong to urinary system disease,because of the high morbidity and mortality,can affects male health seriously.CDK13 belongs to the cyclin-dependent kinase family,play an important role in many biological processes and is involved in the development of drug resistance in various mechanisms.Studies have shown that CDK13 is closely related to various diseases such as congenital heart disease,pancreatic cancer,and ovarian cancer.E2F5 is a member of the E2 F transcription factor family,which plays a vital role in controlling the cell cycle and inhibiting tumor proteins.Studies have shown that E2F5 is closely related to diseases such as chronic obstructive pulmonary disease,breast cancer,liver cancer.At the same time,E2F5 abnormal expression in prostate cancer is associated with Proliferation of prostate cancer cells.However,there is no report about whether CDK13 interacts with E2F5 and what mechanism is happen in prostate cancer.Earlier experiments found that both of them are elevated in prostate cancer.This study aims to explore the mechanisms by which both are involved in prostate cancer,and lay the foundation for the development of new tumor markers and new drugs.Objective: To study the mechanism of the interaction between CDK13 and E2F5 to regulate the proliferation of prostate cancerMethods:1.PCa and Benign Prostatic Hyperplasia clinical specimens were collected,both tissue RNA,protein and pathological sections were extracted.Analyze the different expression of CDK13.2.Perform in vitro experiments to verify the differential expression of CDK13 in four PCa cells(LNCa P,PC3,22RV1,and DU145)and BPH cells(RWPE-1).Proliferation and apoptosis experiments were performed in PCa cells after overexpression or knockdown of CDK13.3.Finding the molecular chaperone(E2F5)of CDK13 by co-immunoprecipitation coupled with mass spectrometry(Co IP-MS)and microarray detection.4.To find out whether connection and co-localization relationship between CDK13 and E2F5 through co-immunoprecipitation experiments,adjacent linking technology(PLA)and immunofluorescence experiments.5.Verify the differential expression of E2F5 in PCa and BPH,compare the results with the TCGA database.Explore whether a correlation between CDK13 and E2F5.6.PCa cells were transfected with vector as NC sg RNA,CDK13 sg RNA,oe E2F5,and sh E2F5,respectively.To overexpress CDK13 or knock down E2F5,perform Western blot experiments,colony formation experiments,cell proliferation experiments,and co-immunoprecipitation experiments to further verify the relationship between the two.Results:1.CDK13 expression is up-regulated in PCa tissue.60 tissue samples detected by q RT-PCR and Westren blot,it was found that the expression of CDK13 in PCa was significantly higher than BPH tissues.2.Knockdown of CDK13 inhibits PCa cell proliferation and promotes its apoptosis.The expression of CDK13 in PC3 and 22RV1 was significantly higher than RWPE-1.Overexpression of CDK13 promoted the proliferation and inhibits its apoptosis of PCa cells,while the knockdown was the opposite.3.In PCa cells,CDK13 interacts with E2F5.Co-immunoprecipitation experiments confirmed the strong interaction between CDK13 and E2F5,ortho-linking technology confirmed the direct binding between the two,and immunofluorescence experiments confirmed the colocalization of the two in PCa tissue.4.E2F5 is highly expressed in PCa tissue.Through q RT-PCR and Western blot experiments,found that the levels of E2F5 m RNA and protein in PCa were higher than BPH.After analysis,there was a positive correlation between E2F5 m RNA and CDK13 m RNA expression in PCa tissues,and the TCGA database confirmed this positive correlation.5.CDK13 and E2F5 interact together to promote PCa cell proliferation.Transfection of CDK13 sg RNA in PCa cells to activate CDK13 transcription can increase the expression of CDK13 and E2F5 in PCa cells,reduce the level of cyclin-dependent kinase inhibitor p21 protein,and promote PCa cell proliferation and inhibits its apoptosis.Transfection of sh E2F5 expressing plasmid to knock down E2F5 in PCa cells can significantly reduce the expression levels of E2F5 and CDK13 and increase the protein level of p21.Conclusions:1.The expression of CDK13 and E2F5 in PCa tissues was significantly up-regulated and positively correlated.2.Knocking down CDK13 can inhibit PCa cell proliferation and promote its apoptosis.3.There is an interaction between CDK13 and E2F5 in PCa cells,and the interaction between CDK13 and E2F5 promotes PCa cell proliferation and inhibits its apoptosis.