TSA联合HSV-1对C6鼠胶质瘤细胞株细胞增殖、细胞凋亡的影响

目的探讨曲古抑菌素A(TSA)联合Ⅰ型单纯疱疹病毒(HSV-1)对C6鼠胶质瘤细胞株细胞增殖、细胞凋亡的影响。方法体外培养C6胶质瘤细胞,设对照组(加入等体积培养基)、TSA组(加入0.5×10^-3μmol/L TSA处理)、HSV-1(加入10 MOI HSV-1处理)及TSA+HSV-1组(加入0.5×10^-3μmol/L TSA和10 MOI HSV-1处理)共4组,每组设5个复孔。CCK-8法检测细胞增殖活性;流式细胞仪检测细胞凋亡率;RT-PCR、免疫印迹法检测血管内皮细胞生长因子(VEGF)的mRNA和蛋白表达水平。结果作用48、72 h,与对照组相比,TSA组、HSV-1组、TSA+HSV-1组C6细胞增殖活性显著降低(P<0.01),细胞凋亡率明显增高(P<0.05),VEGF mRNA和蛋白表达水平明显降低(P<0.05);而且,TSA+HSV-1组明显优于TSA组和HSV-1组(P<0.05)。结论TSA联合HSV-1对体外培养的C6细胞可产生协同或叠加杀伤作用,抑制VEGF表达可能是作用机制之一。

Effects of TSA combined with HSV-1 on cell proliferation and apoptosis of C6 glioma cells

Objective To explore the effect of trichostatin A (TSA) combined with herpes simplex virus type 1 (HSV-1) on the cell proliferation and apoptosis of C6 glioma cells. Methods C6 glioma cells were cultured in vitro, and then randomly divided into four groups, i.e., control group (treatment with equal volume of culture medium), TSA group (treatment with 0.5 × 10-3 μmol/L TSA), HSV-1 group (treatment with 10 MOI HSV-1) and TSA+HSV-1 group (treatment with 0.5 × 10-3 μmol/L TSA and 10 MOI HSV-1). CCK-8 method was used to detect cell proliferation activity. Flow cytometry was used to detect the apoptosis rate. RT-PCR and Western blot were used to detect the mRNA and protein expression of vascular endothelial growth factor (VEGF), respectively. Results Compared with the control group, the proliferation rates of C6 cells were significantly reduced (P<0.01), the apoptosis rates were significantly increased (P<0.05), and the expression levels of VEGF mRNA and protein were significantly reduced (P<0.05) in the TSA group, HSV-1 group, and TSA+HSV-1 group. Moreover, the effect of TSA+HSV-1 group was significantly better than the TSA group and HSV-1 group (P<0.05). Conclusion TSA combined with HSV-1 can produce synergistic killing effects on cultured C6 cells, which may be related to the inhibition of VEGF