端粒酶反义寡聚脱氧核苷酸对MGC-803胃癌细胞的影响  

The Effects of Telomerase Antisense Oligodeoxy Nucleotides on MGC-803 Cells

作　　者：孙大鹏[1] 许德顺[1] 韩冠英[1] 罗喜刚[1] 

机构地区：[1]辽宁医学院,辽宁锦州121001

出　　处：《辽宁医学院学报》2007年第2期6-9,共4页Journal of Liaoning Medical University (LNMU) Bimonthly

摘　　要：目的探讨端粒酶反义寡聚脱氧核苷酸(antisense oligodeoxy nucleotides,ASODN)对MGC-803胃癌细胞端粒酶活性及细胞生长的影响。方法将实验分为正常对照组、端粒酶正义寡聚脱氧核苷酸(sense oligodeoxy nucleotides,SODN)组和不同剂量的ASODN组;脂质体介导的ASODN和SODN作用于MGC-803细胞后,分别采用四甲基偶氮唑蓝比色法(MTT)、酶联免疫吸附法(ELISA)、吖啶橙染色法、流式细胞术检测细胞的体外增殖、端粒酶活性、细胞形态、细胞凋亡和细胞周期的改变。结果转染48h后,终浓度为1、3及5μmol/L的ASODN对MGC-803细胞端粒酶活性及细胞的增殖均有抑制作用,与正常对照组比较差异显著(P<0.01),并呈一定剂量依赖性;流式细胞仪检测到凋亡峰,细胞被阻滞在G1/S期;吖啶橙染色后光镜下观察发现有典型的凋亡小体。结论以端粒酶RNA模板区为靶点的ASODN明显抑制MGC-803胃癌细胞的增殖,其机制可能是通过降低细胞的端粒酶活性而诱发细胞的凋亡,ASODN对胃癌的治疗具有重要价值。Objective This paper has studied the effects of telomerase antisense oligodeoxy nucleotides （ASODN） on telomerase activity and proliferation of MGC - 803 gastric carcinoma cell. Methods MGC - 803 cell was transfected by ASODN, sense oligodeoxy nucleotides （SODN） mediated by Lipotap Liposomal Transfection Reagen. The experiments were classified into normal group, ASODN groups at varied concentrations and SODN group. The proliferation activity of MGC - 803 cell line was determined by using methyl thiazolyl tetrazolium assay. The telomerase activity was determined by using enzyme - linked immunosorbent assay. The cell morphology was observed by fluorescence microscope stained with acridine orange. How cytometry was adopted to examine apoptotic rate and cell cycle. Results 1, 3 and 5μmol/L ASODN significantly inhibited the growth of MGC - 803 cell and the telomerase activity from 48h after transfection. The effect was in dose dependent and sequence specific manner. The apoptotic peak was detected with FCM and cells were stayed in G1/S stage. Typical apoptotic morphologic feature was discovered under light microscope. Conelusions ASODN strongly inhibits the proliferation of MGC - 803 gastric carcinoma cells by inhibiting telomerase activity which induces cell apoptosis. ASODN might be a new solution to gastric carcinoma therapy.

关 键 词：端粒酶反义寡聚脱氧核苷酸 MGC-803胃癌细胞 端粒酶 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]