hTERT,siRNA抑制胆囊癌细胞端粒酶活性及增殖运动探究.docVIP

hTERT,siRNA抑制胆囊癌细胞端粒酶活性及增殖运动探究 　 作者：丁昂　童赛雄　冯平　秦新裕 【摘要】 　　［目的］ 探讨hTERT,siRNA对GBC,SD端粒酶活性、琥珀酸脱氢酶（SDH）活性、侵袭、hTERT mRNA、hTERT蛋白的作用及其相关机制。［方法］在质粒pGCsi,H1/GFP,hTERT干扰后，采用端粒酶TRAP、半定量RT,PCR和Western blot方法检测GBC,SD端粒酶活性、hTERT基因和β,catenin基因mRNA、蛋白质表达水平。四甲基偶氮唑盐（MTT）光吸收法检测SDH活性，运用Transwell小室了解癌细胞侵袭情况。［结果］ pGCsi,H1/GFP,hTERT对GBC,SD端粒酶活性、SDH活性、侵袭、hTERT mRNA、hTERT蛋白均有抑制作用并随浓度升高而增强。pGCsi,H1/negative对上述指标无明显抑制作用，与pGCsi,H1/GFP,hTERT抑制作用比较差异有统计学意义（Plt;0.01）。［结论］ pGCsi,H1/GFP,hTERT通过降低hTERT mRNA及蛋白表达降低，进而抑制端粒酶、SDH活性及其侵袭力。 【关键词】 胆囊肿瘤　RNA干扰　基因　蛋白质　hTERT　β,catenin 　　Inhibitory Effect of hTERT,siRNA on Telomerase Activity Cell Proliferation and Migration in Gallbladder Cancer Cells 2.Huaian Forth Hospital of Suzhou Medical College, Huaian 223300, China）Abstract： ［Purpose］ To explore the of hTERT,siRNA effect on GBC,SD cell telomerase activity，SDH activity， invasion efficiency, hTERT mRNA and hTERT protein and their mechanisms. ［Methods］ Telomerase activity，mRNA and protein of hTERT and β,catenin in GBC,SD were detected by telomerase TRAP，RT,PCR and Western blot techniques，respectively; SDH activity, by MTT; and invasion assay by Transwell chamber after GBC,SD cells were interfered in pGCsi,H1/GFP,hTERT vector. ［Results］ pGCsi,H1/GFP,hTERT could inhibit GBC,SD cell telomerase activity，SDH activity，and invasion efficiency，expression of hTERT mRNA and protein. The inhibition effect was correlated with increase of pGCsi,H1/GFP,hTERT vector concentrations. pGCsi,H1/negative vector had no inhibition on above indexes. A statistically significant difference existed between pGCsi,H1/GFP,hTERT vector and pGCsi,H1/negative vector (Plt;0.01). ［Conclusion］ pGCsi,H1/GFP,hTERT can inhibit telomerase activity，SDH activity and invasion by down regulating the expression of mRNA and protein of hTERT. Key words: gallbladder neoplasms; RNA interference; gene; protein; hTERT; β,catenin β,catenin与胆囊的恶性病变密切相关，端粒酶、hTERT、β,catenin在人胆囊癌细胞系呈阳性表达，其中，hTERT在胆囊癌中阳性表达率为84.1%，它们可提示胆囊癌细胞的生物学特性，影响胆囊癌细胞的增殖、运动和侵袭力[2，3]，因此，