Effects And Mechanism Of TIPE On Proliferation And Differentiation Of Renal Cancer Cells

Objective: Renal cell carcinoma(RCC)is the most common kidney malignancy.The global incidence of renal cell cancer is increasing annually.Advanced RCC is easy to cause systemic multiple metastases with poor prognosis.Early diagnosis and successful partial or total nephrectomy timely can save the lives of patients,but due to its etiology and pathogenesis is still not clear,plus the vast majority of early patients without symptoms,clinical extremely easy to misdiagnosis.Studies have shown that the discovery of new targeted molecules contributes to the early diagnosis and treatment of tumors.Tumor necrosis factor-induced protein 8(TIPE)is an anti-apoptotic oncogene molecule associated with cell proliferation,migration and tumor growth,but its role in RCC remains unclear.This study explored the effect of TIPE on RCC proliferation and migration and analyzed the possible mechanism.Methods: 769-P renal carcinoma cell line was studied.Lentiviral transfection with TNFAIP8(TIPE)-specific shRNAs was used to establish stable TIPE knock down 769-P cell lines.Two groups were divided,that is blank control group and TIPE-shRNA group,according to the treatment.TIPE mRNA expression was detected by RT-PCR.Cell proliferation and viability were assessed by CCK-8 assay Cell cycle was examined by flow cytometry.Expression levels of TIPE,DcR3,ERK1 and ERK2 were detected by Western blot.Results: 1.The size of the electrophoresis band detected by RT-PCR was consistent with that of the designed TIPE gene primer fragment.Compared with the blank control group(1.10�0.09),the expression level of TIPE mRNA in tipe-shrna group decreased significantly(0.51�0.06),and the difference between groups was significant(P<0.05).TIPE protein expression levels in the two groups were(1.78�0.12)and(1.23�.14),respectively.TIPE protein expression was significantly inhibited in the tipe-shrna group compared with the blank control group(P<0.05),and the transfection was successful.2.Silence TIPE gene expression of cell proliferation after dynamic OD values on day 1,2,3,4,0.125,0.215,0.180,0.390,and the control of cell proliferation activity OD values on day 1,2,3,4,0.150,0.243,0.345,0.580,the more similar between the two groups was statistically significant(P<0.05),and longer duration of transfection,TIPE shRNA-769-P cell proliferation ability gradually reduce;After transfection for 48 hours,it was significantly lower than the blank control group,and the difference was statistically significant(P<0.05).Cell migration counts of 769-p cells in tipe-shrna group(308�19)were significantly lower than those in control group(525�23),and the difference was statistically significant(P<0.05).In the blank control group and the tipe-shrna group,the proportion of cells in S phase was(78�10)% and(61�8)%,respectively.The proportion of cells in S phase in the blank control group was significantly higher than that in the tipe-shrna group,and the difference was statistically significant(P<0.05).3.DcR3 and erk1/2 proteins were expressed in both the blank control group and the tipe-shrna group,and the expression level in the blank control group was significantly higher than that in the tipe-shrna group,there is statistically significant differences(P<0.05).Correlation analysis also found that TIPE expression level was positively correlated with DcR3(r=0.733,P<0.05)and ERK1(r=0.590,P<0.05).And its expression level was not correlated with ERK2(P>0.05).Conclusions: TIPE protein was the key target to regulate the proliferation,differentiation and migration of renal carcinoma cells,and DcR3 and ERK pathways were also involved.