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
| 作者: |
1刘春梅,
2丁依玲
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: |
丁依玲
Email: dylcsu@yahoo.com.cn |
| DOI: | 10.3978/j.issn.2095-6959.2013.02.002 |
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。
关键词：
热休克蛋白70；反义寡核苷酸；宫颈癌；HeLa 细胞；增殖；凋亡； 化学治疗敏感性
(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
CorrespondingAuthor: DING Yiling Email: dylcsu@yahoo.com.cn
DOI: 10.3978/j.issn.2095-6959.2013.02.002
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.
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.