端粒酶反义基因对紫杉醇诱导原代胃癌细胞凋亡的增敏作用  

Study on effect of apoptosis induced by telomerase antisense oligodeoxynucleotides and TAXOL in primary human gastric cancer cells

作　　者：崔京远[1] 马红[1] 崔秋娟[1] 周东风[1] 

机构地区：[1]山东省青岛市立医院普外科,山东青岛266011

出　　处：《中国现代普通外科进展》2005年第4期206-208,共3页Chinese Journal of Current Advances in General Surgery

摘　　要：目的:探讨端粒酶反义寡核苷酸(PSASODN)与紫杉醇(TAXOL)联合应用对原代胃癌细胞凋亡作用的影响。方法:常规组织块培养法进行胃癌细胞原代纯化培养,取第3代对数生长期细胞进行实验。各组在培养24h及48h分别加入相同剂量的培养液;终浓度:3μM的PSASODN,3μM的NASODN,1.0μM的TAXOL。作用24h后再分别在PSASODN组及NASODN组加入终浓度为1.0μM的TAXOL;分别于培养后24,48,72,96h收集各组细胞。以台盼蓝拒染法计算各组细胞生长抑制率,观察PSASODN联合TAXOL对原代胃癌细胞生长的影响;流式细胞学观察细胞凋亡率及细胞周期变化。结果:终浓度为3μM的PSASODN作用于原代胃癌细胞24h后加入TAXOL1.0μM,能明显抑制胃癌细胞增殖,同时流式细胞学检测到凋亡峰,细胞受阻于G2/M期,作用48,72,96h的凋亡细胞百分率(34.8%,44.6%及50.6%)明显高于TAXOL组及NASODN+TAXOL组,差异有统计学意义(P<0.05),其作用呈时间依赖性及序列特异性。结论:以端粒酶RNA模板区为靶点的PSASODN可促进TAXOL诱导的胃癌细胞凋亡,对胃癌具有重要治疗价值。Objective：To study the effeot of apoptosis Induced by telomerase antisense ollgodeoxynucleotides（PS-ASODN） and TAXOL in primary human gastric cancer Cells. Methods： Primary Gastric cancer cells were purified by tissue culture method. The third generation Cells were used in this experiment.After incubation for 24 hours and 48 hours,cells were treated with PS-ASODN of 3μM, N-ASODN of 3μM are TAXOL of 1.0μM respectively.After incubation for 24 hours,TAXOL of 1.0μM was added to group PS-ASODN and N-ASODN.Cells were continued to incubate for 48,72 and 96 hours.inhibiting rate was determined by trypan blue.Flow cytometry was adopted to examine apoptotic rate and cell cycle.Results：Treatment with TAXOL after 24 hours of exposure of cells to 3μM PS-ASODN inhibited significantly the growth of primary human gastric cancer cells.The apoptotic peak was detected with FCM and cells were blocked in G2/M stage.The percentage of apoptotic cells treated with TAXOL from 48 hours to 96 hours after 24 hours of exposure to PS-ASODN（34.8%,44.6% and 50.6%）was higher than that of cells treated with N-ASODN plus TAXOL or TAXOL alone（P〈0.05）.The effect was in time dependent and sequence specifice manner.Conclusion：PS-ASODN targeted hTR could increase the TAXOL-induced apoptosis of primary gastric cancer cells.Therefore it may be a new target to gastric carcinoma therapy.

关 键 词：端粒 末端转移酶 寡核苷酸类 反义 紫杉醇 胃肿瘤 细胞凋亡 

分 类 号：R735.2[医药卫生—肿瘤]