Molecular Mechanism By Which NCAPD3 Activates AKT Through STAT3 And PI3K To Promote The Occurrence And Development Of Prostate Cancer

Prostate cancer is a common malignant tumor in male genitourinary system.Its incidence rate and mortality rate are increasing year by year.Incidence rate in Europe and America is second only to lung cancer.With the improvement of the economic level and the changes of people's living standard,eating habits and environment,the incidence rate of prostate cancer is increasing year by year in China.In the early stage,the treatment of prostate cancer is mainly surgery combined with radiotherapy and chemotherapy.In recent years,although the level of basic research and clinical application of prostate cancer continues to improve,the therapeutic effect of prostate cancer is still not ideal.Because the etiology of prostate cancer is very complex,and its specific pathogenesis and molecular mechanism are still unclear,more in-depth and detailed basic research is urgently needed to provide theoretical basis and treatment ideas for the clinical treatment of prostate cancer.NCAPD3 is an important component of condensation protein complex II.Its main function in cells is to regulate the condensation and stabilization of chromatin during mitosis.At present,the research on NCAPD3 is mainly based on the influence of condensation protein complex II on the process of chromatin condensation during mitosis.According to our previous experimental results,NCAPD3,as a downstream target gene of androgen receptor,plays an important role in the occurrence and development of prostate cancer.However,the relationship between NCAPD3 and the development of prostate cancer and its molecular mechanism have not been reported,which need to be further explored.AKT,also known as protein kinase B(PKB),is a serine / threonine kinase.It is often abnormally activated by high phosphorylation modification in a variety of cancers,and participates in a variety of physiological processes of cancer cells,such as proliferation,survival,metabolic reprogramming,invasion / metastasis,inhibition of autophagy and aging,thus promoting cancer progression.Previous experiments in our laboratory proved that NCAPD3 can promote AKT phosphorylation,but the specific molecular mechanism is not clear.Therefore,based on the existing research results,this project further explored the molecular mechanism of NCAPD3 regulating AKT phosphorylation and its related molecular signaling pathway from the following aspects:Firstly,the expression of NCAPD3 in prostate cancer tissues and adjacent normal tissues was analyzed by TCGA database.The results showed that compared with other cancers,the expression of NCAPD3 in prostate cancer tissues was more significant;the expression level of NCAPD3 in prostate cancer tissues was significantly higher than that in adjacent normal tissues;patients with high expression of NCAPD3 had a worse prognosis.These results suggest that NCAPD3 plays an important role in the development of prostate cancer.Secondly,NCAPD3 was knocked down by siRNA in LNCa P and overexpressed in DU145.The phosphorylation of AKT(Thr308 and Ser473)was detected by Western blot.The results showed that there was a positive correlation between NCAPD3 and p-AKT(Thr308/Ser473),which was further verified in DU145-NCAPD3 cells.Then,Transwell assay and scratch assay were used to detect the migration of prostate cancer cells in DU145 cells overexpressing NCAPD3,and cell proliferation was detected by colony formation assay.The results showed that overexpression of NCAPD3 significantly enhanced the migration and proliferation of prostate cancer cells,while LY294002 significantly reversed the inhibition of PI3 K activity.These results suggest that NCAPD3 affects the migration and proliferation of prostate cancer cells through PI3 K / AKT pathway.Western blot was used to detect the levels of STAT3 / EZH2 / NSD2 / m TORC2 and STAT3 / EZH2 / p16 / e EF1A2 / PI4 KBeta / PDK1 pathway.The results showed that overexpression of NCAPD3 significantly increased the levels of STAT3,EZH2,NSD2,e EF1A2,PI4 KBeta,and down regulated the levels of p16.In the cells with knockdown of NCAPD3,the results were just the opposite.Then,overexpression of NCAPD3 in DU145 cells and knockdown of NCAPD3 in LNCa P cells were used to detect the related factors in JAK2 / PI3 K / PDK1 pathway.It was found that overexpression of NCAPD3 significantly increased the protein levels of JAK2 and p-PI3K;knockdown of NCAPD3 resulted in the opposite results.These results indicate that NCAPD3 phosphorylates AKT(Thr308 and Ser473)and activates AKT through the above pathways,thus promoting the progression of cancer.In conclusion,NCAPD3 can activate downstream NSD2,EEF1A2 and PDK1 by regulating JAK2 / PI3 K and STAT3 / EZH2 pathways,and then phosphorylate AKT(Thr308 and Ser473)to activate AKT,thus promoting the occurrence and development of prostate cancer.Through this study,we explored the specific molecular mechanism of NCAPD3 promoting prostate cancer,provided experimental basis for the further improvement of the molecular etiology of prostate cancer,and provided new drug targets for the treatment of prostate cancer in the future.