Effects Of SPOCK1 Gene On Invasion And Proliferation Of Bladder Cancer Cells

Background:The incidence and mortality of bladder cancer are increasing year by year,which is the most common malignant tumor of urinary system in china.Nowadays,transurethral resection of bladder tumor,radical cystectomy and urinary diversion,postoperative radiotherapy and chemotherapy are still the most important and effective methods for the treatment of bladder cancer.But even after surgery,adjuvant chemotherapy,bladder cancer treatment is still not ideal,the reason attributed to bladder cancer recurrence and metastasis.Relapse and metastasis are the main causes of treatment failure and death.The mechanism of the occurrence and development of bladder cancer is not clear,it is necessary to study its pathogenesis and look for effective therapeutic targets.The extracellular matrix glycoprotein encoded by SPOCK1(Sparc/osteonectin cwcv and kazal-like domains proteoglycan 1)gene has been proved to be closely related to the proliferation,adhesion and metastasis of tumor cells.High expression of SPOCK1 gene has been found in bladder cancer tissues,and the high expression of SPOCK1 is associated with poor prognosis of urothelial carcinoma.To objective to investigate the effect of SPOCK1 gene on the development of bladder cancer,and to provide experimental evidence for the mechanism of bladder cancer,we observed the changes of invasion and proliferation by Knockdowning of SPOCK1 gene in bladder cancer cells.Purpose:1、To verify the expression of SPOCK1 gene in bladder cancer cells.2 、 We further investigated the effect of SPOCK1 gene on invasion and proliferation of bladder cancer cells In vitro cell experiment.Method:1.QRT-PCR and Western Blot were used to detect the expression of the target gene SPOCK1 in human bladder epithelial immortalized cells(SV-HUC-1),bladder cancer cell line 5637 and bladder cancer cell line T24.2.SPOCK1 small interfering RNA fragments(sh SPCOK1)were constructed and transfected into bladder cancer cell line 5637 by liposome-mediated transfection.The protein of SPOCK1 was detected by Western Blot method.The experimental group was divided into blank control group(Control cells group),Scrambled si RNA group and experimental group(SPOCK1 si RNA group).3.The expression of SPOCK1 gene was decreased by RNAi technology in bladder cancer cell line 5637.The changes of invasion and proliferation of bladder cancer cells were detected by Wound scratch assay、 Transwell assay and CCK-8assay.Result:1.The results of q RT-PCR and Western Blot showed overexpression of SPOCK1 related m RNA and protein in bladder cancer cell lines and the expression of spock1 was higher in bladder cancer cell line 5637 than that of T24.2.Compared with other interference sequences,the interference effect of SPOCK1-si RNA-2 sequence is the best(P<0.05).3.The invasion and proliferation of bladder cancer cells were significantly inhibited by wound scratch assay,Transwell assay and CCK-8 assay,after the expression of SPOCK1 gene in bladder cancer cell line 5637 was decreased by RNAi technology.Conclusion:1.SPOCK1 gene expression is increased in bladder cancer cells.2.The expression of SPOCK1 gene promotes the invasion and proliferation of bladder cancer cells,which may be a promising target for gene therapy of bladder cancer.