Apigenin/GCV对C_6-TK细胞株的杀伤效应及作用机制

The killing effect and its mechanism about C6 lines transfected with TK gene after treated with Apigenin/GCV after in vitro

目的探讨Apigenin/GCV对体外培养鼠胶质瘤C6细胞的杀伤效应及作用机制。方法对含有导入pcDNA3-TK质粒的C6细胞进行培养、传代:通过MTT法、TUNEL实验法及光镜技术检测GCV/Apigenin对检测不同组别C6细胞C6-TK细胞株的杀伤效应。结果给GCV+Apigenin后:转染组、空载体组和对照组中两两比较:转染组的细胞增殖水平显著低于空载体组和对照组(P<0.05);相对未处理组,用10μmol/LApigenin预先处理6h后,GCV对C6混合细胞的杀伤效应显著增强(P<0.01)。结论缝隙连接功能上调剂Apigenin可显著提高HSV-TK/GCV肿瘤自杀基因系统的"旁观者效应"及显著促凋亡作用。GCV的作用效能对细胞间通讯功能有一定的依赖性。

Objective The study aimed to investigate the killing effect to C6 lines and the bystander effect induced by Apigenin/GCV after transfected with TK gene in vitro in order to gain a result for the gene therapy of glioma.Methods C6 lines transfected with HSV-TK plasmid DNA in vitro were cultured and proliferated.The techniques of MTT,TUNEL and Fluorescent microscope were used to determine apoptosis,cell-cycle and proliferation in difference group.Results The growth of Co lines were inhibited evidently after treated with apigenin.The cells showed characters of apoptosis and differentiation observed by light microscopy and FCM.The rates of apoptosis increased after treated with GCV and more markedly increased after treated with apigenin/GCV in six houses in C6-TK＋ group than C6-TK-and C6 lines（P〈0.01）.Conclusion Apigenin,which was reported as Gap Junctiunal intercellular communication up-regulator can inhibit the growth,proliferation and differentiation of C6 lines and induce its apoptosis significantly.The effection of GCV was relaed to the function of the gap Junctiunal intercellular communication of C6 lines and its changes modulated by chemical agent in some degree.