RNA干扰阻断COX-2基因表达对食管鳞状上皮癌EC109细胞株增生和凋亡的影响

首都医科大学学报 ›› 2008, Vol. 29 ›› Issue (3): 295-299.

RNA干扰阻断COX-2基因表达对食管鳞状上皮癌EC109细胞株增生和凋亡的影响

张莉¹, 张澍田¹, 于中麟¹, 李鹏¹, 宗晔¹, 许彩民², 华方圆², 韩兵社²

1. 首都医科大学附属北京友谊医院消化科;2. 中国医学科学院中国协和医科大学基础医学研究所国家分子生物学实验室

收稿日期:2007-05-31 修回日期:1900-01-01 出版日期:2008-06-24 发布日期:2008-06-24
通讯作者: 张澍田

The Effect of RNA Interference Silencing Cyclooxygenase-2 Gene on Proliferation and Apoptosis of Esophageal Squamous Carcinomas EC109 Cell

Zhang Li¹, Zhang Shutian¹, Yu Zhonglin¹, Li Peng¹, Zong Ye¹, Xu Caimin², Hua Fangyuan², Han Bingshe²

1. Department of Digestive Diseases, Beijing Friendship Hospital, Capital Medical University;2. National Laboratory of MedicalMolecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College

Received:2007-05-31 Revised:1900-01-01 Online:2008-06-24 Published:2008-06-24

摘要/Abstract

摘要： 目的研究COX-2基因与食管鳞癌细胞EC109增生和凋亡的关系,观察阿司匹林对食管鳞癌细胞EC109增生和凋亡的影响,探索基因治疗和药物抑制肿瘤的机制.方法采用RNA干扰的方法,用荧光显微镜观察RNA干扰的转染效率,用Westernblot方法分析RNA干扰的效果,用噻唑兰法定量检测RNA干扰和阿司匹林对食管鳞癌细胞增生的影响,用流式细胞仪检测RNA干扰和阿司匹林对食管鳞癌细胞凋亡的影响.结果 Westernblot方法证实,RNA干扰后EC109细胞COX-2表达下降至对照组的40%.MTT法检测结果显示,用COX-2基因干扰后,EC109细胞生长抑制作用增强,COX-2基因干扰和阿司匹林联合作用后,肿瘤细胞的生长抑制作用进一步增强.流式细胞仪检测结果显示,母本细胞组和RNA干扰的阴性对照组凋亡率较低,COX-2基因干扰后,促凋亡作用增加.COX-2基因干扰后再给予阿司匹林,促凋亡作用进一步增加.结论 RNA干扰特异性抑制细胞COX-2蛋白的表达后,抑制食管鳞癌细胞株EC109的生长增生,增强凋亡作用,阿司匹林和RNA干扰对肿瘤细胞的增生和凋亡可产生协同效果.

关键词: 食管鳞状上皮癌, 环氧化酶-2, 阿司匹林, 凋亡, RNA干扰

Abstract: Objective To study the relationship between cox-2 gene and the proliferation as well as apoptosis of esophageal squamous carcinoma EC109 cell at the gene level;also by studying the effects of aspirin(non-specific cooxygenase-2 inhibitor) on proliferation and apoptosis of esophageal squamous carcinoma EC109 cell,to investigate the mechanism of gene and medical therapies for esophageal squamous carcinoma.Methods The RNA interference method was used in this study.Short interfere RNA was synthesized targeting Cox-2 Gene.The expression of COX-2 gene in esophageal squamous cell carcinoma EC109 was specially inhibited by RNAi technique.The efficiency of RNAi technique was detected with fluorescence microscopy.The effect of RNAi technique was detected with Western blot.Cell growth was tested with MTT reduction assay.The cell apoptosis was detected with flow cytometry.EC109 growth was tested with MTT reduction assay.Results Compared with the control group,the expression of COX-2 gene in EC109 had a 40% decrease.The OD value of MTT was 0.85±0.05 in mother cell group,the OD value of MTT was 0.75±0.06 in negative control group labeld by FITC,the OD value of MTT was 0.48±0.05 in transfection group of COX-2 siRNA,the OD value of MTT was 0.28±0.04 in Aspinin combined with RNAi group.After RNAi,the effect of growth inhibition was increased in EC109 cell compared with the control groups.The growth inhibition of Aspinin combined with RNAi group was stronger than that of RNAi group.The rate of apoptosis was 1.35% in mother cell group,the rate of apoptosis was 1.72% in negative control group labeld by FITC,the rate of apoptosis was 4.33% in transfection group of COX-2 siRNA,the rate of apoptosis was 11.66% in Aspinin combined with RNAi group.After RNAi,the rate of apoptosis increased in EC109 cell compared with the control groups.The rate of apoptosis of aspinin conbined with RNAi group was stronger than that of RNAi group.Conclusion The RNAi techniques can specifically inhibit the expression of COX-2 gene.The cell proliferation has been suppressed and the cell apoptosis is stronger in EC109 after COX-2 gene has been specifically inhibited.The combined administration of aspirin and RNAi has a synergic effect on the tumor cell proliferation and apoptosis.

Key words: esophageal squamous cell carcinoma, cooxygenase-2, aspirin, apoptosis, RNA interference

中图分类号:

R735.1

张莉;张澍田;于中麟;李鹏;宗晔;许彩民;华方圆;韩兵社. RNA干扰阻断COX-2基因表达对食管鳞状上皮癌EC109细胞株增生和凋亡的影响[J]. 首都医科大学学报, 2008, 29(3): 295-299.

Zhang Li;Zhang Shutian;Yu Zhonglin;Li Peng;Zong Ye;Xu Caimin;Hua Fangyuan;Han Bingshe. The Effect of RNA Interference Silencing Cyclooxygenase-2 Gene on Proliferation and Apoptosis of Esophageal Squamous Carcinomas EC109 Cell[J]. Journal of Capital Medical University, 2008, 29(3): 295-299.

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