Bcl-Xl基因反义寡核苷酸诱导食管癌细胞凋亡的研究  

Induction of apoptosis in human esophageal cancer cells by Bcl-Xl antisense oligodeoxynucleotide

作　　者：张蕾[1] 郭克民[2] 温洪涛[3] 唐琳[4] 张云汉[4] 

机构地区：[1]河南省肿瘤病理重点实验室郑州大学临床医学博士后流动站,450052 [2]郑州大学医学院法医教研室 [3]郑州大学第一附属医院消化内科 [4]河南省肿瘤病理重点实验室郑州大学第一附属医院病理科

出　　处：《中华检验医学杂志》2005年第9期953-956,共4页Chinese Journal of Laboratory Medicine

摘　　要：目的探讨BclXl基因反义寡核苷酸(ASODN)对食管癌细胞株EC9706增殖和凋亡的影响。方法用阳离子脂质体介导bclxL基因ASODN转染EC9706细胞后,采用四甲基偶氮唑盐光吸收法(MTT法)检测EC9706细胞的增殖抑制率;逆转录聚合酶链反应和蛋白印迹杂交检测BclXlmRNA和蛋白表达水平;吖啶橙荧光染色和流式细胞术定量检测EC9706细胞的凋亡率。结果MTT检测显示,BclXlASODN可显著抑制EC9706细胞的增殖(P<0.01),而且其对细胞增殖的抑制作用具有剂量依赖性;BclXlASODN对BclXlmRNA表达抑制率为57.29%。用吖啶橙荧光染色法和流式细胞检测ASODN组凋亡率(分别为31.13%±5.75%和30.5%,与细胞对照组、空白对照组及无关序列寡核苷酸组比较,差异均有统计学意义(均P<0.05)。结论BclXl基因ASODN可有效抑制EC9706细胞的增殖,通过ASODN作用可下调BclXl基因表达,并显著促进EC9706细胞凋亡;BclXl基因有望成为食管癌基因治疗的新靶点。Objective To investigate the induction of proliferation and apoptosis in esophageal cancer cell line EC9706 by Bcl-Xl antisense oligodeoxynucleotide （ ASODN ）. Methods Bcl-XL ASODN was transfected into EC9706 following cationic liposome. The proliferation inhibitory rate of EC9706 was assessed by MTT. Expressions of Bcl-Xl were detected by RT-PCR and Western Blot. Cell apoptosis were observed by acridine orange （AO） fluorescent staining and flow cytometry, respectively. Results The proliferation of EC9706 was decreased significantly by Bcl-Xl ASODN in dose-dependent manner （P 〈 0. 01 ）, In the group of ASODN, the expression inhibitory rate of Bcl-X1 mRNA was 57. 29% and the apeptosis rates detected by AO fluorescent staining and flow cytometry were 31.13% ± 5.75% and 30. 5%, respectively. There were significant difference as compared with the cell control group.the vacuity control group and the non-sense oligonucleotide group, respectively （P 〈 0.05 ）. Conclusion Bcl-Xl ASODN can effectively inhibit the proliferation of EC9706 cells, and significantly induce apoptnsis of EC9706 cells by suppressing the expression of Bcl-Xl. The results suggest that Bcl-Xl may be a promising target for gene therapy in human esophageal carcinoma.

关 键 词：食管肿瘤 寡核苷酸类 反义 BCL-XL基因 基因反义寡核苷酸 食管癌细胞株 癌细胞凋亡 EC9706细胞 逆转录-聚合酶链反应 吖啶橙荧光染色 ASODN 

分 类 号：R735.1[医药卫生—肿瘤]