论著 Original Article

热休克蛋白70反义脱氧寡核苷酸对宫颈癌HeLa细胞生长和凋亡及化学治疗敏感性的影响

Published at: 2013年第33卷第2期

刘春梅 1 , 丁依玲 2
1 河南省宁陵县人民医院妇产科,河南 宁陵 476700
2 中南大学湘雅二医院妇产科,长沙410011
通讯作者 依玲 丁 Email: dylcsu@yahoo.com.cn
DOI: 10.3978/j.issn.2095-6959.10.3978/j.issn.2095-6959.2013.02.002
基金:

摘要

目的:探讨反义脱氧寡核苷酸封闭HSP70基因对体外宫颈癌 HeLa 细胞增殖、凋亡及化疗敏感性的影响。方法:1)将体外培养的宫颈癌 HeLa 细胞分为正常对照组(Ctrl组)、反义寡核苷酸处理组(AS组)、正义寡核苷酸处理组(S组)、随机寡核苷酸处理组(R组),每组各5例,分别转染体外培养的宫颈癌 HeLa 细胞,采用Western免疫印迹检测各组细胞HSP70蛋白表达。2)将顺铂处理体外培养的宫颈癌 HeLa 细胞分为正常对照组(Ctrl组)、单纯顺铂处理组(Cis组)、反义寡核苷酸+顺铂处理组(AS +Cis组)、正义寡核苷酸+顺铂处理组(S +Cis组)、随机寡核苷酸+顺铂处理组 (R +Cis组),每组各8例。采用四甲基偶氮唑蓝光吸收法(methyl thiazolyl tetrazolium,MTT) 法检测 HeLa 细胞的生长抑制率; 流式细胞术检测 HeLa 细胞的凋亡率。结果:1)Ctrl组、AS组、S组、R组HSP70灰度比值分别为1.365±0.187,0.379±0.134,1.403±0.163和1.410±0.158,转染AS组HSP70灰度比值明显下降(P<0.01);2)AS+Cis组生长抑制率和凋亡率明显高于其它各组,差异有统计学意义(P<0.05),S+Cis组和R+Cis组细胞生长抑制率和凋亡率与Cis组相比差异无统计学意义(P>0.05)。结论:HSP70反义脱氧寡核苷酸通过下调HeLa 细胞中HSP70蛋白表达,抑制细胞增殖并诱导细胞凋亡。HSP70反义脱氧寡核苷酸与顺铂联合具有协同作用,能提高细胞对顺铂化学治疗的敏感性。


Effect of heat shock protein 70 antisense oligodeoxynucleotides on growth, apoptosis and chemosensitivity for human cervical cancer HeLa cells

Abstract

Objective: To explore the effect of heat shock protein 70 (HSP70) antisense oligodeoxynucleotides on proliferation, apoptosis and chemosensitivity for HeLa cells induced by cisplatin. Methods: 1)HeLa cells were transfected by antisense, sense or random oligonucleotides. The cells were divided into a control group(Ctrl group, n=5), a antisense oligodeoxynucleotides treatment group (AS group, n=5), a sense oligodeoxynucleotides treatment group (S group, n=5) and a random oligonucleotides treatment group (R group, n=5). The expression of heat shock proteins 70 (HSP70) in the cells from each group was detected. 2) The HeLa cells were treated by cisplatin and were divided into a control group (control group, n=8), a cisplatin treatment group (Cis group, n=8), a antisense oligodeoxynucleotides + cisplatin treatment group (AS +Cis group, n=8), a sense oligodeoxynucleotides + cisplatin treatment group (S +Cis group, n=8) and a random oligonucleotides + cisplatin treatment group (R +Cis group, n=8). The growth inhibitory rate of HeLa cells was determined by methyl thiazolyl tetrazolium (MTT) assay. The apoptosis rate was detected by flow cytometry (FCM). Results: 1)The optical density of HSP70 band in the Ctrl group, the AS group, the S group, and the R group were (1.365 ±0.187) , (0.379 ±0.134), (1.403 ±0.163) and (1.410±0.158), respectively, which was significantly decreased in the AS group (P<0.01). 2)Compared with the control group, the growth inhibitory rate and the apoptosis rate of HeLa cells were significantly increased only in AS+Cis group (P<0.05). Conclusion: HSP70 antisense oligodeoxynucleotides can effectively inhibit the proliferation of HeLa cells and induce its apoptosis through down-regulation of HSP70 expression. HSP70 antisense oligodeoxynucleotides can increase the chemosensitivity of tumor to cisplatin treatment.


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引用本文: 春梅 刘, 依玲 丁. 热休克蛋白70反义脱氧寡核苷酸对宫颈癌HeLa细胞生长和凋亡及化学治疗敏感性的影响[J]. 临床与病理杂志, 2013, 33(2): 98-104.
Cite this article as: LIU Chunmei, DING Yiling . Effect of heat shock protein 70 antisense oligodeoxynucleotides on growth, apoptosis and chemosensitivity for human cervical cancer HeLa cells[J]. Journal of Clinical and Pathological Research, 2013, 33(2): 98-104.