中文摘要：

目的将p CMV-Cx43c DNA重组质粒转染入大鼠脑胶质瘤C6细胞,上调C6细胞Cx43表达,进而探讨C6细胞Cx43过表达对其增殖能力的影响及机制。方法通过脂质体将p CMV-Cx43c DNA重组质粒转入C6细胞,使C6细胞过表达Cx43,并用G418筛选出稳定过表达Cx43的C6细胞;测定细胞倍增时间和软琼脂集落形成实验,检测各组细胞的增殖程度;分别用ERK1/2特异性阻断剂PD98059（30μmol·L^-1）、p38MAPK特异性阻断剂SB20219（10μmol·L^-1）处理各组细胞,Western blot检测各组细胞Cx43、p-Cx43、pERK1/2和p-p38MAPK的表达;MTT比色法检测各组细胞活性。结果成功将p CMV-Cx43c DNA重组质粒转入C6细胞,并筛选出稳定转染Cx43基因的C6细胞;测定细胞倍增时间和软琼脂集落形成实验表明C6-Cx43组比C6组、C6-p CMV组细胞增殖速度减慢,细胞集落数明显减少（P〈0.01）;ERK1/2、p38MAPK特异性阻断剂处理细胞后,Western blot检测发现C6-Cx43＋PD98059组、C6-Cx43＋SB202190组较C6-Cx43组Cx43表达升高（P〈0.01）、pCx43（P〈0.05）表达下调。结论阻断ERK1/2、p38MAPK通路,导致Cx43蛋白表达升高,从而抑制C6细胞的增殖。

英文摘要：

Aim To explore the effect of Cx43 over-expression on proliferation of C6 cells and its mechanisms by transfecting p CMV-Cx43 c DNA plasmid into C6 cells.Methods p CMV-Cx43 c DNA plasmid was transfected into C6 cells by liposome to up-regulate the expression of Cx43,and C6 cells with over-expression of Cx43 was stably cloned by using G418.Determination of cell doubling time and soft agar colony formation assay to detect the degree of cell proliferation.The cells were treated with ERK1/2 specific blocker PD98059（30 μmol · L^-1） and p38 MAPK specific blocker SB202190（10 μmol · L^-1） respectively,the expression of Cx43,p-Cx43,p-ERK1/2 and p-p38 MAPK of each group were detected by Western blot,and the activity of each group was detected by MTT Assay.Results p CMV-Cx43 c DNA plasmid was transfected into C6 cells successfully.Cell lines with over-expression Cx43（C6-Cx43） or empty vector（C6-p CMV） were stably selected by using G418.Determination of cell doubling time and soft agar colony formation experiments showed that the proliferative rate and the colony number of C6-Cx43 group were significantly decreased,compared with that of C6 group and C6-p CMV group（P〈0.01）;ERK1/2,p38 MAPK specific blockers were treated with each group,Western blot showed that the expression of Cx43 protein was increased（P〈0.01）,while p-Cx43 protein was decreased（P〈0.05） in C6-Cx43 ＋ PD98059 group and C6-Cx43 ＋SB202190 group,compared with that of C6-Cx43 group.Conclusion Cx43 may decrease the proliferation of glioma cells through ERK1/2,p38 MAPK pathways.