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

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(10μmol/L)、紫杉醇(10 nmol/L)+SAHA(10μmol/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IC 50。RT-PCR法检测各组HeLa细胞中抑癌基因p27 m RNA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IC 50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 m RNA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。

OBJECTIVE:To investigate the inhibitory effect of paclitaxel and SAHA on human cervical cancercells.METHODS:Human cervical cancer(HeLa)cells were treated with paclitaxel(10nmol/L),SAHA(10μmol/L),or paclitaxel(10nmol/L)+SAHA(10μmol/L)for24hours or48hours.The inhibitory rates of HeLa cells were determinedby MTT assay.p27mRNA levels were determined by RT-PCR assay,protein levels of Ac-H4were identified by Westernblot and IC of paclitaxel was calculated by SPSS16.0.RESULTS:Results from the MTT assay show that the inhibition50rates from exposure to the combination of paclitaxel and SAHA were significantly higher than those from the singletreatment groups(P<0.01).Similarly,RT-PCR assay show that the p27mRNA levels from the combined treatments weresignificantly higher than from single treatments(P<0.01).From combined treatments,the inhibitory rates were54.27%±4.02%with24h treatment and77.02%±3.86%with48h treatment,the mRNA level was10.601±0.673,the protein level of Ac-H4was1.282±0.033,and IC was reduced significantly.CONCLUSION:SAHA in combination with50paclitaxel significantly inhibited HeLa cell proliferation by enhancing the level of histone acetylation and inducing theexpression of tumor suppressor genes in vitro.