摘要
目的探讨T-cadherin分子表达抑制胶质母细胞瘤C6细胞增殖的分子机制。方法利用脂质体将pcDNA3和pcDNA3.T-cadherin表达质粒转染C6细胞,筛选出表达T-cadherin的C6克隆3、4和5以及转染空载体后不表达T-cadherin的C6克隆1和2,以Western Bolt检测各克隆的p21和p27的表达;同时,以pcDNA3和pcDNA3.T-cadherin表达质粒分别转染野生型(p21+/+)和p21缺失突变型(p21-/-)成纤维细胞,研究T-cadherin分子介导的细胞增殖抑制是否依赖细胞周期蛋白p21的表达。结果转染pcDNA3.T-cadherin质粒后表达T-cadherin分子的C6克隆3、4和5的p21表达水平明显升高,而p27的表达与C6细胞是否表达T-cadherin无关。另外,转染pcDNA3.T-cadherin质粒的野生型表达p21的成纤维细胞的增殖显著受抑,而转染pcDNA3.T-cadherin质粒对p21缺失突变的成纤维细胞的增殖不产生抑制作用。结论T-cadherin分子抑制胶质母细胞瘤C6细胞的增殖与p21表达密切相关,且T-cadherin分子对成纤维细胞的增殖抑制作用依赖于p21的表达。
Objective To study the molecular mechanism of growth suppression mediated by T-cadherin expression in C6 glioma cells. Methods C6 cells were transfected with pcDNA3 and pCDNA3. T-cadherin using lipofectin. The T-cadherin-expressing C6 clone 3, 4, 5 and vector-transfected C6 clone 1, 2 were screened. The expression levels of p21 and p27 levels were detected by Western blot. Wild-type fibroblast (p21 + / + ) and p21 mutated fibroblast (p21-/-) were transfected with pcDNA3 and pCD-NA3. T-eadherin to study whether growth suppression mediated by T-cadherin expression depended on the p21 expression. Results The p21 expression levels in T-cadherin-expressing C6 clone 3, 4 and 5 were aignificandy increased, but the p27 levels were not related to T-cadherin expression. The cell growth of wild type fibroblast (p21 + / + ) was significantly suppressed after transfected with pCDNA3. T-cad- herin, but the cell growth of p21 mutated fibroblast (p21-/-) was affected by the transfection of pCD- NA3. T-eadherin. Conclusion Growth suppression mediated by T-cadherin expression in C6 cells is closely related to the p21 expression. Moreover, the suppression mediated by T-cadherin in fibroblast dependa on the p21 expression.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第11期1304-1305,共2页
Chinese Journal of Experimental Surgery