Effect of lentivirus-mediated MFN2 overexpression on the proliferation of renal carcinoma cells

Abstract

Objective To investigate the effect of mitofusion-2 (MFN2) on the proliferation of renal carcinoma cells and to explore its molecular mechanism. Methods ACHN and A498 cells were infected with Lenti - MFN2 cells harboring MFN2 coding sequence (experimental group) , lentivirus containing green fluorescent protein gene (Lenti-GFP) was used as the control group. After transfection, real-time quantitative PCR (qRT-PCR) and Western blotting were used to determine the mRNA and protein expression of MFN2 and phosphorylation of Ras-Raf1-ERK1/2 signaling pathway. The cell cycle was detected by flow cytometry. MTS assay and colony formation assay were used to detect changes in cell viability and proliferation. Results In the control group and experimental group, the expression of MFN2 mRNA in ACHN cells were 1.04±0.29 vs 4.11±0.78 (P<0.001) , respectively; in A498 cells, 1.02±0.24 vs 4.84±1.49 (P=0.002) , respectively. In the experimental group, the expression of MFN2 protein was significantly increased, the phosphorylation level in Ras-Raf1-ERK1/2 signaling pathway significantly decreased, the cell cycle inhibited and the viability and proliferation ability of renal carcinoma cells significantly lowered. Conclusion MFN2 may inhibit the cell cycle process by inhibiting protein phosphorylation in the Ras-Raf1-ERK1/2 signaling pathway, resulting in decreased viability and proliferation of renal cell carcinoma cells. Key words: Mitochondrial fusion protein 2; Renal neoplasms; Cell cycle; Cell proliferation