摘要
目的观察载体介导的RNA干扰(RNAi)技术对人喉鳞癌Hep-2细胞增殖及其血管内皮生长因子C(VEGF-C)mRNA表达的影响。方法选择针对人特异性VEGF-C的小RNAi(siRNA)靶序列mRNA,分别设计、合成含有19 nt干扰序列的寡核苷酸(同时随机组合出一条对照序列),将两两互补的寡核苷酸经退火后所形成的序列克隆到pSuper-neo载体中构建重组体。脂质体法转染重组体至喉鳞癌Hep-2细胞,经G418筛选获得阳性克隆。用流式细胞仪检测细胞周期,RT-PCR检测其VEGF-C mRNA。结果转染VEGF-C靶向siRNA载体的Hep-2细胞生长减慢,S期细胞显著减少,VEGF-C表达水平明显下降。结论VEGF-C靶向RNAi重组体可显著抑制Hep-2细胞增殖及其VEGF-C mRNA的表达。
Objective To observe the inhibitory effect of RNAi on cell proliferation and expression of vascular endothelial growth factor C （VEGF-C） mRNA in laryngeal carcinoma cells. Methods A vector for small interference RNA （siRNA） targeting VEGF-C gene was constructed, introduced into laryngeal carcinoma Hep-2 cells and the positive clones were screened. The cell proliferation was observed by FCM and the VEGF-C mRNA of Hep-2 were detected by RT-PCR. Result The VEGF-C mRNA was blocked in Hep-2, with S phase cell decreased. Conclusion The small RNAi targeting VEGF-C can inhibit the cell proliferation and the expression of VEGF-C mRNA of Hep-2.
出处
《山东医药》
CAS
北大核心
2008年第18期19-20,共2页
Shandong Medical Journal
关键词
RNA干扰
血管内皮生长因子C
喉肿瘤
细胞增殖
RNA interference
vascular endothelial growth factor C
laryngeal neoplasms
cell proliferation
