Research On The Influence Of C-Src Gene On Biological Characteristics Of Renal Cell Carcinoma And Its Clinical Significance

[Objective]The proto-oncogen c-Src is closely relevant with tumor development. In previous research we discovered the nuclear import of CXCR4 promoted the metastasis of renal cancer via the interaction with c-Src, and in this study we intend to use the gene knockdown technique to research the influence of the lenti-virus c-Src shRNA on renal cell carcinoma and the clinical significance, complete the research on the CXCL12/CXCR4 biological axis signal pathway, and provide the theoretical basis for the clinical decision on the targeted therapy of renal cell carcinoma.[Methods]This study verified the research objective via the following 3 aspects:1. Preparing the lenti-virus induced c-Src shRNA and using RT-PCR and Western blot to verify the gene knockdown effect on the level of mRNA and protein.2. Using in vitro CCK8 cell proliferation assay, Transwel and flow cytometry to verify the changes of proliferation activity, migration and invasion, cell cycle and apoptosisof of renal cancer cell line after the knockdown of c-Src.3. Using the immunohistochemistry and tissue microarray to detect the expression of c-Src gene in rencal cancer tissue, and investigating the clinical significance of c-Src gene combining the analysis of clinical index and prognosis.[Results]1. Successfully screened a c-Src shRNA lenti-virus with knockdown efficiency of 73%, and verified it can significantly suppress the mRNA and protein expression of c-Src gene in A498-EGFP and A498-EGFP-CXCR4 cell line, as well as the mRNA and protein expression of CXCR4 gene.2. Comparing with negative control, the cell proliferation activity and invasion and migration ability decreased in stable transfected A498-EGFP-SRC shRNA and A498-EGFP-CXCR4-SRC shRNA renal cancer cell line, the cell cycle arrested in S phase, and the apoptosis was not significant.3. In tissue sample, the expression of c-Src correlated with the patients’ prognosis. The survival of patients with different expression of c-Src were significant different, and the prognosis of patient with high expression of c-Src was worse.[Conclusion]After the c-Src gene knockdown the mRNA and protein expression of c-Src gene decreased, combining with the decreased mRNA and protein expression of CXCR4 gene, which further verified the nuclear import of CXCR4 may interact with c-Src to promote tumor cell metastasis. In vitro research menifested the c-Src gene knockdown can decrease the biological activity of renal cancer cell. The survival of renal cancer patients with high expression of c-Src in tissue microarray was significant shortened. Thus, the c-Src gene may become new target of renal cancer therapy.