RNAi沉默c-Met基因对人脑胶质瘤U251细胞增殖及侵袭能力的影响

目的  探讨RNAi沉默c-Met基因对人脑胶质瘤U251细胞增殖及侵袭能力的影响。方法  分别构建3种靶向c-Met基因的短发卡RNA（shRNA）序列，以脂质体转染的方式导入人脑胶质瘤U251细胞，并筛选出稳定表达的细胞株。采用PCR和Western blot检测c-Met-shRNA转染后U251细胞c-Met的表达情况，以筛选出可有效沉默c-Met基因的shRNA。c-Met基因沉默后，采用MTT法和流式细胞术检测U521细胞增殖情况，采用Transwell法和细胞划痕实验检测U251细胞侵袭和迁移情况，采用裸鼠荷瘤实验研究U251细胞体内成瘤性。结果  成功构建可有效沉默c-Met基因的c-Met-shRNA1。沉默c-Met后，c-Met mRNA以及蛋白的表达均显著下降，差异有统计学意义（P <0.05）；而且可明显抑制U251细胞的增殖、侵袭、迁移以及体内成瘤能力，差异有统计学意义（P <0.05）。结论  RNAi沉默c-Met基因可有效抑制人脑胶质瘤U251细胞的增殖、侵袭、迁移以及体内成瘤能力。

Effect of RNAi silencing c-Met gene on proliferation and invasion ability of human glioma U251 cell

Objective To investigate the effect of RNAi silencing c-Met gene on the proliferation and invasion ability of human glioma U251 cell. Methods Three kinds of short hairpin RNA (shRNA) targeting c-Met gene were constructed. All of the shRNAs were transfected into U251 cell by liposome transfection, in order to screen out the stable expression cell line. The expression of c-Met in U251 cell was detected by PCR and Western Blot, in order to screen out the effective shRNA. The proliferation of U251 cell was detected by MTT assay and flow cytometry. The invasion and migration of U251 cell was detected by transwell assay and cell scratch. Results The shRNA targeting c-Met gene was successful constructed. The expression of c-Met was significantly decreased after c-Met gene was silenced, the difference was statistically significant (P < 0.05). The proliferation, invasion, migration and tumorigenicity of U251 cell was inhibited after silencing c-Met gene, the difference was statistically significant (P < 0.05). Conclusions RNAi silencing c-Met gene can effectively inhibit the proliferation, invasion, migration and tumorigenicity of human glioma U251 cell.