RNAi沉默Cyr61基因对喉癌细胞增殖、凋亡及侵袭能力的影响  

The influence of Cyr61 siRNA on cell proliferation,apoptosis and invasion in human laryngeal cancer cell line Hep-2

作　　者：陈向军[1] 余滋中[1] 李国义[1] 

机构地区：[1]郧阳医学院附属太和医院耳鼻喉科,湖北十堰442000

出　　处：《现代肿瘤医学》2009年第12期2263-2266,共4页Journal of Modern Oncology

摘　　要：目的:观察RNA干扰技术沉默Cyr61基因表达对人喉癌Hep-2细胞生物学行为的影响。方法:针对Cyr61mRNA的序列设计合成小干扰RNA(siRNA)的DNA模板,构建pRNAT-Cyr61重组质粒,转染人喉癌Hep-2细胞;RT-PCR和Western blot检测其对Hep-2细胞内源性Cyr61表达的影响;四甲基偶氮唑蓝(MTT)法观察Hep-2细胞体外增殖活性变化;Transwell小室法检测Hep-2细胞体外侵袭能力的改变;流式细胞仪检测Hep-2细胞凋亡。结果:pRNAT-Cyr61重组质粒在mRNA及蛋白水平分别显著抑制Cyr61基因表达,抑制率分别最高达到74.62%和78.43%;Hep-2细胞的体外生长抑制率最高达68.22%,侵袭细胞数下降至(44.00±2.35)个;Hep-2细胞凋亡率最高达52.98%。结论:pRNAT-Cyr61可抑制Cyr61在喉癌Hep-2细胞中的表达,并抑制细胞的增殖活性和侵袭能力,促进细胞凋亡。Objective：To explore the relationship between the expression of Cyr61 and the biological behaviors of human laryngeal cancer Hep -2 cells by decreasing Cyr61 gene expression by RNA interference. Methods： One pair of DNA template coding was synthesized against Hep -2 to reconstruct pRNA - Cy161, which was transfected into Hep -2 cells. The Cyr61 expression in Hep -2 cells were transfeeted with pRNAT - Cyr61, and it was detected by RT - PCR. Western blot was used to observe the expression of Cyr61 ;MTT,Transwell and FCM were used to observe the biological behaviors of Hep -2 cells. Results： RT - PCR and Western blot analyses demonstrated that pRNAT - Cyr61could significantly inhibit the expression of Cyr61 to 74.62% and 78.43% in Hep - 2 cells; MTT, Transwell and FCM results showed that pRNAT - Cyr61 can significantly inhibit the proliferation by 68.22% and reduce the invasive cell numbers to（44.00_＋2.35 ）. The occurrence of apoptosis increased significantly to 52.98% in Hep-2 cells in vitro. Conclusion： pRNAT - Cyr61 could significantly inhibit the expression of Cyr61 ,suppress the growth and invasion of Hep -2 cells and induce it apoptosis.

关 键 词：CYR61 RNA干扰 HEP-2 喉癌 

分 类 号：R730[医药卫生—肿瘤] R739.65[医药卫生—临床医学]