慢病毒介导的RASSF1A表达抑制小细胞肺癌H446细胞的生长~

RASSFIA expression mediated by lentivirus inhibits growth of small-cell lung cancer cell line H446

目的:探讨Ras相关结构域家族1A(Ras-association domain family 1A,RASSF1A)抑制小细胞肺癌细胞生长的机制。方法:构建含有RASSFlA基因的慢病毒表达载体,包装成病毒颗粒,感染H446小细胞肺癌细胞。MTT测细胞生长曲线;流式细胞术测细胞周期;real-time PCR和Western blotting分别检测细胞周期相关蛋白mRNA和蛋白水平的表达。结果:成功获取稳定表达RASSF1A的H446细胞(RASSF1A-H446)。RASSF1A-H446细胞生长缓慢,细胞周期阻滞在G_1期。Real-time PCR和Western blotting结果显示在RASSF1A-H446细胞中,p21和p27的表达明显升高,E2F1的表达明显降低。结论:RASSF1A通过升高p21、p27和降低E2F1的表达,抑制H446细胞的生长。

AIM： To explore the inhibitory effect of Ras-association domain family 1A （ RASSF1A） on the small-cell lung cancer cell growth .METHODS：The lentiviral expression vector containing RASSF1A gene was constructed and used to infect the small-cell lung cell line H446.The growth curve and cell cycle were detected by MTT assay and flow cytometry.The mRNA and protein levels of cell cycle-associated proteins were determined by real-time PCR and Western blotting.RESULTS：We obtained the H446 cells in which RASSF1A was stably expressed （named RASSF1A-H446）. Compared with normal cell group and negative cell group , RASSF1A inhibited the proliferation of H446 cells, and arrested H446 cells in G1 phase.The expression of p21 and p27 was significantly increased , and E2F1 was significantly decreased in RASSF1A-H446 cells.CONCLUSION：RASSF1A inhibits the H446 cell growth by increasing the expressions of p 21 and p27, and decreasing the expression of E 2F1.