RNAi技术特异抑制HPV18E6基因表达的研究  

Specific suppression of HPV18 E6 oncoprotein by short interference RNA in Hela cervical carcinoma cells

作　　者：聂妹芳[1] 陶光实[1] 刘凤英[1] 林秋华[1] 

机构地区：[1]中南大学湘雅二医院妇产科,湖南长沙410011

出　　处：《中国现代医学杂志》2009年第3期370-374,共5页China Journal of Modern Medicine

摘　　要：目的探讨利用RNAi技术研究靶向抑制宫颈癌细胞株Hela中HPV18E6的可行性。方法根据哺乳动物SiRNA设计原则要求,利用T7RNA聚合酶体外合成针对HPV18E6、E7的SiRNA,用2%的琼脂糖电泳鉴定、脂质体转染Hela细胞,分4组,A组:HPV18E6的SiRNA干预组;B组:HPV18E7的SiRNA干预组;C:HPV18E6、E7的SiRNA干预组;D:基因组文库任意SiRNA为对照组。于转染48、72、96h后,用免疫组化法测定HPV18E6蛋白的表达。结果干预组A和C转染后48、72h,HPV18E6的阳性表达明显减少,以72h为明显,96h后表达无明显减少。而干预B组和对照D组的HPV18E6的阳性表达无明显改变。结论靶向HPV18E6的SiRNA能特异瞬时抑制靶基因E6的表达,该方法将为宫颈癌预防和治疗提供新的策略。[Objective] By RNAi, we investigate whether HPV18 E6 SiRNA targeted Hela cells cervical carcinoma can inhibit the expressions of HPV18 E6 oncoprotein. [Methods] For SiRNA studies in mammalian cells, we synthesize HPV18 E6 and E7 SiRNAs in vitro using T7 RNA polymerase. The products were checked by running on 2% agarose gel. Then Hela cells were grown in standard conditions and transfecting with lipofeetarnineTM. The cells were divided into four groups, A group： HPV18 E6 SiRNA; B group： HPV18 E7 SiRNA; C group： HPV18 E6＋E7 SiRNA; D group： control. After transfection 48, 72 and 96 hours, we investigated the expressions of HPV18 E6 in Hela cells of different groups by S-P immunohistoehemical methods. [Results] The expressions of HPV18 E6 in group A and C were significant decreased after transfeeted 48 hours, 72 hours, especially after transfeetion 72 hours, but have no difference after transfected 96 hours. In group B and D, the expressions of HPV18 E6 is similar to those before transfected with SiRNA E6 and E7. [Conclusions] Targeted HPVE6 SiRNA can specific inhibit the expressions of HPV18 E6 oncoprotein and it may be provided a new methods of cervical carcinoma gene therapy.

关 键 词：宫颈癌 HPV RNAI 免疫组化 

分 类 号：R737.33[医药卫生—肿瘤]