末端转移酶_

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

http://www.100md.com 《中国普通现代外科进展》 2005年第4期

末端转移酶,,端粒，末端转移酶·寡核苷酸类，反义·紫杉醇·胃肿瘤·细胞凋亡,1材料和方法,2结果,3讨论,参考文献

     【摘要】目的：探讨端粒酶反义寡核苷酸(PSASODN)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PSASODN,3μM的NASODN，10μM的TAXOL。作用24h后再分别在PSASODN组及NASODN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PSASODN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PSASODN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NASODN+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PSASODN可促进TAXOL诱导的胃癌细胞凋亡，对胃癌具有重要治疗价值。

    【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

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

    CUI Jingyuan,MA Hong,CUI Qiujuan,ZHOU Dongfeng

    Department of General Surgery,Qingdao Municipal Hospital(Qingdao 266011,China)

    【ABSTRACT】Objective:To study the effect of apoptosis induced by telomerase antisense oligodeoxynucleotides(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 for24 hours and 48 hours,cells were treated with PSASODN of 3μM,NASODN of 3μM and 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. ......

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