生存素基因沉默增强人舌鳞状细胞癌Tca8113对顺铂敏感性的实验  

Silencing of survivin gene enhances chemosensitivity of human tongue cancer cell line Tca8113 to cisplatin

作　　者：许建辉[1] 潘朝斌[1] 黄洪章[1] 张彬[1] 王建广[1] 张磊涛[1] 

机构地区：[1]中山大学附属第二医院口腔颌面外科,广州510120

出　　处：《中华口腔医学杂志》2007年第5期280-283,共4页Chinese Journal of Stomatology

基　　金：国家自然科学基金(30271423)

摘　　要：目的探讨靶向生存素短发夹状 RNA(short hairpin RNA,shRNA)对人舌鳞状细胞癌Tca8113细胞株生存素基因表达、凋亡及其对顺铂、5-氟尿嘧啶(5-Fu)敏感性的影响。方法脂质体介导的生存素 shRNA 表达载体转染 Tca8113细胞,未转染、转染脂质体及错配 shRNA 载体的细胞作为对照。采用 RT-PCR 和 Western blot 分别检测生存素 mRNA 和蛋白的表达,流式细胞仪分析细胞的凋亡率,MTT 法测定抗癌药物的敏感性。结果生存素 shRNA 表达质粒转染后,与3个对照组相比较,生存素 mRNA 和蛋白表达水平明显下降,细胞凋亡率的上升呈时间依赖性,48h 可达37.9%。生存素 shRNA 能显著增加顺铂的敏感性,与未转染组比较其 IC_(50)值下降约4/5(P<0.01),但对5-Fu的作用不明显。结论靶向生存素的 shRNA 可有效抑制 Tca8113细胞生存素 mRNA 和蛋白的表达,同时增加了顺铂的化疗敏感性。Objective To investigate the effects of survivin short hairpin RNA （shRNA） on survivin expression, cell apoptosis, and cheomosensitivity of humantongue cancer cell Tca8113 to cisplatin. Methods Survivin-directed shRNA plasmid vector was delivered into Tca8113 cells with lipofectamineTM 2000 reagent. Survivin expression was detected with the reverse transcription-polymerase chain reaction （ RTPCR） and Western blotting. Flow cytometry was used to examine cell apoptosis, and the sensitivity to anticancer agents was evaluated by methyl thiazolyl tetrazolium （MTTY） assay. Results After survivin shRNA vector transfection in Tca8113 cells, the expression of mRNA/protein declined significantly, and the apoptotic rate increased in time-dependent manner up to 37.9% at 43 hours. RNAi-mediated survivin reduction selectively inhibited growth and enhanced chemosensitivity of cisplatin but not of 5-fluorouracil. Conclusions Survivin shRNA could inhibit the expression of survivin mRNA and it＇s protein and euhance the chemosensitivity of cisplatin.

关 键 词：癌 鳞状细胞 抗药性 肿瘤 顺铂 氟尿嘧啶 

分 类 号：R739.8[医药卫生—肿瘤]