RNA干扰抑制Skp2表达对人喉癌细胞系Hep-2的影响

Effect of RNA Interference Inhibiting Skp2 Expression on Human Laryngeal Carcinoma Cell Line Hep-2

[目的]研究RNA干扰抑制Skp2表达对喉癌鳞状细胞p27表达、细胞增殖和凋亡的作用。[方法]利用脂质体将重组质粒转染Hep-2细胞,用慢病毒系统建立干扰Skp2基团的稳定细胞株;荧光实时定量PCR和Western blot法检测转染细胞株中Skp2和p27m RNA及其蛋白表达;采用MTT法和流式细胞仪检测转染细胞的增殖和凋亡情况。[结果]通过慢病毒siRNA技术,Skp2可持续稳定抑制Hep2喉癌细胞,siRNA可诱导抑制Skp2,增加p27表达,降低细胞增殖,增加喉癌细胞的凋亡。[结论]靶向Skp2基因的si RNA对Hep-2细胞的抑制作用可能是通过上调p27基因水平来实现的,Skp2是基因疗法治疗喉癌的有利靶点。

[Purpose] To investigate the effect of RNA interference Skp2 suppression on p27 expression,cellular proliferation,apoptosis in human laryngeal carcinoma cell line Hep-2. [Methods]Recombinant plasmids were transfected into Hep-2 cell with liposome. To construct the stable cell line interfered Skp2 gene with lentivirus system. Fluorescent realtime quantitated PCR and Western blot were used to detect expression of m RNA and protein Skp2 and p27 in transfected cell lines. MTT and flow cytometer were used to detect the proliferation and apoptosis in transfected cell lines. [Results] Skp2 was stably suppressed in Hep2 laryngeal carcinoma cells by lentivirus siRNA technology. The siRNA-induced suppression of Skp2 increased p27 expression,decreased cellular proliferation and increased apoptosis in human laryngeal carcinoma cells. [Conclusion]The inhibitation effect of si RNA targeting Skp2 on Hep-2 cell are probably derived from the increased expression of p27 gene,Skp2 is a viable target in the gene therapy treatment for human laryngeal carcinoma.