过度表达G蛋白调节子16促进大鼠胶质瘤C6细胞增殖

Promotion of glioma C6 cells proliferation by overexpressed RGS16

目的 探讨 G蛋白调节子 16 (RGS16 )对大鼠胶质瘤C6细胞的生物学特性的影响 .方法 利用脂质体介导法将RGS16基因导入 C6细胞中 ,在倒置显微镜下观察细胞形态变化和贴壁情况 ;3H- Td R法检测 C6细胞在转染不同梯度p CMV5 - RGS16和 p CMV 5质粒后的增殖情况 ;免疫细胞化学法检测转染前后 RGS16蛋白的表达情况 ;流式细胞仪检测转染 p CMV5 - RGS16和 p CMV5质粒 36 h后细胞周期变化和细胞是否有凋亡发生 .结果 转染 p CMV5 - RGS16质粒 2 4 h后 30 %细胞贴壁性降低 ,突起收缩 ,细胞变圆 ;RGS16蛋白表达阳性 ;3H- Td R法检测显示 C6细胞增殖速度与转染p CMV5 - RGS16的量呈正相关 ;细胞周期结果显示 G1期细胞百分数减少 10 % ,而 S期细胞百分数增多 14 % ;未发现RGS16与凋亡有直接关系 .结论 RGS16可能促进 C6细胞的增殖 .

s: AIM To study the effect of RGS16 on the biological characteristics of glioma C6 cells. METHODS pCMV5 RGS16 was transfected into C6 cells by lipofectin. The morphological and adhesive changes of the cells were observed under an inverted microscope. Proliferation of C6 cells was measured by 3H thymidine ( 3H TdR) assay after gradient transfections of pCMV5 RGS16 and pCMV5. Expression of RGS16 was examined by immunocytochemical method both before and after the transfection. Flow cytometry was adopted to measure changes in the fraction number of the cell cycle phase and to detect whether RGS16 could induce apoptosis of C6 cell. RESULTS 24 hours after the transfection of pCMV5 RGS16 approximately 30% of C6 cells grew round and 13% expressed RGS16; 36 h later the positive relationship between the proliferation of C6 cells and the gradient transfections of pCMV5 RGS16 was displayed by 3H TdR assay. Flow cytometry showed that the fraction number of G1 phase of C6 cells reduced by 10% and that of S phase accumulated by 14% and RGS16 could not induce apoptosis of C6 cells. CONCLUSION RGS16 might promote the proliferation of C6 cells.