摘要
目的采用RNA干扰技术探讨生存素(survivin)对人肺腺癌细胞增殖及凋亡的影响及其可能的分子机制。方法采用脂质体介导将pGCsi-sh survivin与pGCsi质粒转染人肺腺癌细胞株A549,经G418筛选获得稳定抑制survivin表达的肺腺癌细胞模型;Western blot检测转染细胞survivin、p21、鼠双微体基因2(MDM2)蛋白表达;MTT比色法检测阳性细胞克隆的增殖活性;流式细胞仪检测各组细胞凋亡。结果成功构建稳定抑制survivin表达的肺腺癌细胞模型。与对照组相比,反义组p21表达及细胞凋亡率显著增加,而MDM2表达及细胞增殖显著降低。结论下调A549细胞survivin表达后,细胞增殖降低而细胞凋亡增加,其作用机制与p21表达增加和MDM2降低有关。
Objective To explore the influence of RNA - interfered survivin on the prolifera- tion and apoptosis of human lung adenocareinoma cells and its possible mechanism. Methods pGCsi -shsurvivin and pGCsi plasmids were transfected into human lung adenocarcinoma A549 cells. The protein expression of survivin, p21 and MDM2 were detected by Western blot. The cell proliferation and apoptosis were observed by MTT assay and flow cytomertry. Results Survivin -inhibited adeno carcinoma cell models were successfully established. Compared with the control group, the expression of p21 and apoptosis rate increased significantly in pGCsi -sh survivin transfected group, while the MDM2 expression and cell proliferation reduced significantly. Conclusion After down regulation of survivin expression of A549 cells, the cells proliferation reduce and cells apoptosis increase, and relat ed mechanism may be associated with increasing expression of p21 and reduction of MDM2.
出处
《实用临床医药杂志》
CAS
2013年第24期45-47,60,共4页
Journal of Clinical Medicine in Practice
基金
江苏省"333工程"培养资金资助项目(200924)
江苏省自然科学基金资助项目(BK2010357)
关键词
RNA干扰
生存素
鼠双微体基因2
细胞凋亡
RNA interference
survivin
murine double minute gene 2
cell apoptosis
