Background & objective: Wild type p53 could induce RGS16 mRNA expression, and RGS16 protein is related with carcinogenesis of glioma. This study was designed to investigate the effects of exogenous RGS16 gene transfection on the growth of rat glioma cell line C6.
Methods: A eukaryotic expression plasmid pIRES2-EGFP-RGS16 was constructed, and transfected into C6 cells. The cells were selected by G418. The expression of enhanced green fluorescent protein (EGFP) was observed under fluorescent microscope, and the expression of RGS16 mRNA and protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. The effects of RGS16 gene on cell cycle progression, cell growth, and proliferation of C6 cells were measured by flow cytometry, cell growth curve, and clonogenic assay in plate.
Results: C6-RGS16 and C6-GFP cells, which separately expressed RGS16 and pIRES2-EGFP vector, were constructed successfully. S phase proportion was significantly higher in C6-RGS16 cells than in C6-GFP and C6 cells (28.5% vs. 18.9% and 14.3%, P<0.05); C6-RGS16 cells grew faster than C6-GFP and C6 cells. The clone formation rate was significantly lower in C6-RGS16 cells than in C6-GFP and C6 cells (12% vs. 25% and 25%, P<0.05).
Conclusion: Exogenous RGS16 gene could promote cell cycle progression of glioma C6 cells from G1 phase to S phase, and accelerate cell growth, but can't increase the clone formation.