The Mechanism Of PRDX3 Involved In The Development And Progression In Clear Cell Renal Cell Carcinoma

AimTo investigate the relationship between the expression of PRDX3(thioredoxin-dependent peroxide reductase)and the development of ccRCC(clear cell renal cell carcinoma),based on the ccRCC tissues and related cell lines and over expression and knockdown techniques.MethodsThe expression of PRDX3 was first verified in 16 cases of ccRCC tissues and adjacent normal tissues.The expression of different proteins in ccRCC and adjacent tissues were analyzed by quantitative proteome technology,and the expression of PRDX3 was verified.The PRDX3 recombinant lentivirus vector and PRDX3 RNAi vector were constructed with the control cell lines.In the present study,according to the PRDX3 level in ccRCC tissues,we established the stable PRDX3 overexpression cell lines and knockdown cell lines in 786-0 cell lines.qPCR and western blot were used to verify the overexpression and knockdown of PRDX3.The growth of tumor cells in those cell lines was detected by CCK-8 test.Interaction proteins with PRDX3 were searched by anti-flag pull-down test combined with LC-MS/MS technique.The interaction between PRDX3 and PRDX1 was preliminarily explored.ResultsThe western blot results showed that PRDX3 was down-regulated in 14 out of 16 ccRCC tissue samples about 1.78 times.Quantitative proteome results showed that PRDX3 was down-regulated in 12 of the 16 samples,which was basically consistent with western blot results.The ratio is 1.89 times which the cutoff value is 1.35 times.Stable PRDX3 overexpression and knockdown cell lines and those control group were successfully established(7860-PRDX3(+)and 7860-PRDX3(-),7860-PRDX3 KN and 7860-PRDX3 NCi).PRDX3 expression in 7860-PRDX3(+)was 2.1 times higher than 7860-PRDX3(-)at mRNA level and 1.8 times at protein level.PRDX3 expression in 7860-PRDX3 KN was 0.48 times lower than 7860-PRDX3 NCi at mRNA level and 0.51 times at protein level.The cell growth rate of 7860-PRDX3(+)cell lines was significantly lower than that of 7860-PRDX3(-).Meanwhile,there was no significant difference in 7860-PRDX3 KN and NCi cell lines.Pull-down results show that PRDX3 may interact with PRDX1 through disulfide bond and the binding sites of those two proteins were identified respectively.ConclusionPRDX3 was down-regulated expression in renal clear cell carcinoma and the interaction with PRDX1 may be involved in the development of tumor.Increasing the expression level of PRDX3 can significantly reduce the growth rate of tumor cells.PRDX3 is involved in the development of renal clear cell carcinoma through interaction with PRDX1.Based on PRDX3,it is possible to develop targeted drugs for treating renal clear cell carcinoma.