The Role And Preliminary Mechanism Of KNTC1 In The Occurrence And Development Of Bladder Cancer

[Objective]To explore the role and possible mechanism of KNTC1 gene in the occurrence and development of bladder cancer.[Methods]The expression level of KNTC1 in bladder urothelial carcinoma tissues and adjacent transitional epithelium tissues of 46 patients and other bladder urothelial carcinoma tissues of 13 patients were detected by immunohistochemistry.The correlation between the expression of KNTC1 and clinical features and overall survival was analyzed.RNA interference was used to knock down the expression of KNTC1 in EJ and T24 bladder cancer cell lines.MTT assay was used to detect cell proliferation,flow cytometry was used to detect apoptotic cells and cell cycle,wound healing assay was used to detect cell migration,and Transwell assay was used to detect cell invasion.The tumorigenesis of T24 bladder cancer cell line with knockdown of KNTC1 expression in nude mice was detected by in vivo tumorigenesis experiment.Protein antibody microarray and Western blot were used to detect the expression of KNTC1 knockdown related proteins,in order to explore its possible regulatory mechanism.[Results]Immunohi stochemi cal results showed that the expression of KNTC1 in bladder cancer tissues was significantly higher than that in adjacent tissues.The expression of KNTC1 was significantly different in different pathological grades(grade Ⅱ and Ⅲ),different T stages(Tis/T1/T2 and T3/T4)and different clinical stages(Ois/1/2 and 3/4).The expression of KNTC1 in higher pathological grades,T stages and clinical stages was higher than that in lower pathological grades,T stages and clinical stages.The high expression of KNTC1 gene was a risk factor for overall survival,and the death risk of high expression was 3.139 times that of low expression.The results of cell function tests showed that compared with the control group,the proliferation of bladder cancer cells in KNTC1 knockdown group was significantly slowed down,the apoptosis was significantly increased,the percentage of G2 phase cells was significantly increased,and the migration and invasion ability was significantly inhibited.Compared with the control group,the tumorigenicity of T24 cells in KNTC1 knockdown group was significantly inhibited.The results of protein antibody microarray showed that the expression levels of Caspase3 and Fas were significantly up-regulated after KNTC1 gene knockdown,and the expression levels of Bcl-2,Bcl-w,CD40,c1AP-2,HSP60,IGF-I,IGF-1sR,Livin,sTNF-R2,TNF-α,TNF-β and XIAP was significantly down regulated.[Conclusion]KNTC1 is highly expressed in bladder cancer,which is significantly correlated with clinicopathological features and prognosis.Knockdown of KNTC1 expression can inhibit the proliferation of bladder cancer cells,increase apoptosis,inhibit cell migration and invasion,and inhibit tumor formation,suggesting that KNTC1 plays an important role in the occurrence and development of bladder cancer,and can be used as a biomarker and potential therapeutic target of bladder cancer.Knockdown of KNTC1 gene can change the expression level of apoptotic signal pathway proteins,suggesting that KNTC1 plays a role by affecting apoptotic signal pathway.