组蛋白去乙酰化酶抑制剂联合紫杉醇抑制宫颈癌细胞增殖的体外实验

doi:10.3969/j.issn.1004-616x.2017.05.003

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (5): 335-339.doi: 10.3969/j.issn.1004-616x.2017.05.003

组蛋白去乙酰化酶抑制剂联合紫杉醇抑制宫颈癌细胞增殖的体外实验

刘好^1,2, 赵滢滢², 金平³, 许改霞⁴, 郭宇然⁴, 张可⁵, 王晓梅^1,2, 张蕾^3,5

1. 深圳大学生命科学与海洋学院, 广东深圳 518060;
2. 深圳大学医学部基础医学院生理学教研室, 广东深圳 518060;
3. 深圳市妇幼保健院妇科, 广东深圳 518048;
4. 深圳大学光电工程学院, 广东深圳 518060;
5. 深圳市罗湖区人民医院妇科, 广东深圳 518020

收稿日期:2017-03-15 修回日期:2017-08-13 出版日期:2017-09-30 发布日期:2017-09-30
通讯作者: 王晓梅,E-mail:xmwang@szu.edu.cn;张蕾,E-mail:1481215091@qq.com E-mail:xmwang@szu.edu.cn;1481215091@qq.com
作者简介:刘好,E-mail:liuhao_35@sina.com。
基金资助:
深圳市科技创新基础研究项目（JCYJ20140415163543959，JCYJ 20140418091413563）

Histone deacetylase inhibitor restrained proliferation of human cervical cancer cells

LIU Hao^1,2, ZHAO Yingying², JIN Ping³, XU Gaixia⁴, GUO Yuran⁴, ZHANG Ke⁵, WANG Xiaomei^1,2, ZHANG Lei^3,5

1. College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060;
2. Department of Physiology, School of Basic Medical Sciences, Shenzhen University Health Sciences Center, Shenzhen 518060;
3. Gynecology of Shenzhen Maternity & Child Healthcare Hospital, Shenzhen 518048;
4. College of Optoelectronic Engineering, Shenzhen University, Shenzhen 518060;
5. Gynecology of Shenzhen Luohu People's Hospital, Shenzhen 518020, Guangdong, China

Received:2017-03-15 Revised:2017-08-13 Online:2017-09-30 Published:2017-09-30

摘要/Abstract

摘要： 目的：研究组蛋白去乙酰化酶抑制剂SAHA联合紫杉醇对宫颈癌HeLa细胞增殖的抑制效果及其机制。方法：设置空白对照组、紫杉醇（10 nmol/L）、SAHA（10 μmol/L）、紫杉醇（10 nmol/L）+SAHA（10 μmol/L）联合组，采用四甲基噻唑蓝（MTT）法检测各组HeLa细胞的生长抑制率，计算紫杉醇对HeLa细胞的IC₅₀。RT-PCR法检测各组HeLa细胞中抑癌基因p27 mRNA的相对表达，Western blot检测HeLa细胞乙酰化组蛋白H4（Ac-H4）的表达。结果：紫杉醇、SAHA、紫杉醇+SAHA联合组处理HeLa细胞24 h的相对抑制率分别为25.93%±5.32%、46.38%±3.66%、54.27%±4.02%，联合组抑制率与紫杉醇组相比明显升高，差异具有统计学意义（P < 0.01）；48 h的抑制率分别为29.12%±3.09%、65.26%±3.03%、77.02%±3.86%，联合组抑制率最强，显著高于SAHA组和紫杉醇组（P < 0.01）。经SAHA预处理后紫杉醇对HeLa细胞的IC₅₀较单独紫杉醇组均显著下降（P < 0.05或P < 0.01）。紫杉醇组、SAHA组、紫杉醇+SAHA联合组细胞中p27 mRNA的相对表达量分别为5.845±0.548、0.978±0.117和10.601±0.673，乙酰化组蛋白H4（Ac-H4）的相对表达分别为0.878±0.068、1.148±0.018、1.282±0.033，联合组中表达量均显著高于SAHA组和紫杉醇组（P均 < 0.01）。结论：SAHA联合紫杉醇在体外能显著抑制宫颈癌细胞HeLa的增殖，增强组蛋白乙酰化水平，诱导抑癌基因的表达。

关键词: SAHA, 紫杉醇, 宫颈癌, HeLa细胞, 细胞增殖

Abstract: OBJECTIVE: To investigate the inhibitory effect of paclitaxel and SAHA on human cervical cancer cells. METHODS: Human cervical cancer (HeLa) cells were treated with paclitaxel (10 nmol/L),SAHA (10 μmol/L),or paclitaxel (10 nmol/L) + SAHA (10 μmol/L) for 24 hours or 48 hours. The inhibitory rates of HeLa cells were determined by MTT assay. p27 mRNA levels were determined by RT-PCR assay,protein levels of Ac-H4 were identified by Western blot and IC₅₀ of paclitaxel was calculated by SPSS 16.0. RESULTS: Results from the MTT assay show that the inhibition rates from exposure to the combination of paclitaxel and SAHA were significantly higher than those from the single treatment groups (P < 0.01). Similarly,RT-PCR assay show that the p27 mRNA levels from the combined treatments were significantly higher than from single treatments (P < 0.01). From combined treatments,the inhibitory rates were 54.27%±4.02% with 24 h treatment and 77.02%±3.86% with 48 h treatment,the mRNA level was 10.601±0.673,the protein level of Ac-H4 was 1.282±0.033,and IC₅₀ was reduced significantly. CONCLUSION: SAHA in combination with paclitaxel significantly inhibited HeLa cell proliferation by enhancing the level of histone acetylation and inducing the expression of tumor suppressor genes in vitro.

Key words: SAHA, paclitaxel, cervical cancers, HeLa cell, cell proliferation

中图分类号:

R730.1

刘好, 赵滢滢, 金平, 许改霞, 郭宇然, 张可, 王晓梅, 张蕾. 组蛋白去乙酰化酶抑制剂联合紫杉醇抑制宫颈癌细胞增殖的体外实验[J]. 癌变·畸变·突变, 2017, 29(5): 335-339.

LIU Hao, ZHAO Yingying, JIN Ping, XU Gaixia, GUO Yuran, ZHANG Ke, WANG Xiaomei, ZHANG Lei. Histone deacetylase inhibitor restrained proliferation of human cervical cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(5): 335-339.

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组蛋白去乙酰化酶抑制剂联合紫杉醇抑制宫颈癌细胞增殖的体外实验

Histone deacetylase inhibitor restrained proliferation of human cervical cancer cells

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