Cx43对脑胶质瘤C6细胞增殖的影响及机制

发布时间：2018-03-25 09:26

本文选题：脑胶质瘤C细胞　切入点：缝隙连接　出处：《中国药理学通报》2017年07期

【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(P0.01);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(P0.01)、pCx43(P0.05)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。
[Abstract]:Objective to transfect the recombinant plasmid of p CMV-Cx43c DNA into C6 cells of rat brain glioma and up-regulate the expression of Cx43 in C6 cells, and to investigate the effect and mechanism of Cx43 overexpression on the proliferation of C6 cells. Methods the recombinant plasmid of p CMV-Cx43c DNA was transfected into C6 cells by liposome. C6 cells were overexpressed Cx43, and C6 cells stably expressed Cx43 were screened by G418. The cell doubling time and soft Agar colony forming test were measured to detect the proliferation degree of the cells in each group. ERK1/2 specific blockers, PD98059(30 渭 mol LPERK-1, p38MAPK-specific blocker, SB20219(10 渭 mol Lamp-1a) were used to detect the expression of Cx43, p-Cx43, pERK1 / 2 and p-p38MAPK, respectively. Results the recombinant plasmid p#en6# DNA was successfully transferred into C6 cells, and the expression of pERK1 / 2 and p-p38MAPK were detected by MTT colorimetric assay. The recombinant plasmid p38 MAPK was transfected into C6 cells successfully, and the expression of pERK1 / 2 and pERK1 / 2 were detected by MTT colorimetry. C6 cells stably transfected with Cx43 gene were screened, and cell multiplication time and soft Agar colony formation test showed that the proliferation rate of C6-Cx43 group was slower than that of C6 C6-p CMV group. The number of cell colonies decreased significantly after treatment with P0.01ERK1 / 2p38MAPK specific blocker. Western blot analysis showed that the expression of C6-Cx43 SB202190 in C6-Cx43 PD98059 group was higher than that in C6-Cx43 group. Conclusion blocking ERK1 / 2 p38 MAPK pathway leads to the increase of Cx43 protein expression and inhibits the proliferation of C6 cells.
【作者单位】：南昌大学药学院药理学教研室;
【基金】：国家自然科学基金资助项目(No 81660014,30760286) 江西省自然科学基金资助项目(No 20142BAB205022)
【分类号】：R739.41

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