摘要
目的探讨应用小干扰核糖核酸(siRNA)干扰组蛋白去乙酰化酶1(HDAC1)的表达,提高人类宫颈癌He La细胞对顺铂(DDP)化疗敏感性的作用与机制。方法合成针对HDAC1的siRNA采用脂质体法转染宫颈癌He La细胞,将其分为siRNA组、阴性siRNA组及对照组。对照组、siRNA组和阴性siRNA组通过DDP处理后分别作为对照+DDP组、siRNA+DDP组和阴性siRNA+DDP组。采用PCR检测HDAC1 mRNA表达,采用流式细胞仪检测细胞凋亡率,采用四甲基偶氮唑蓝(MTT)比色法检测He La细胞对DDP的敏感性。结果与阴性siRNA组及对照组比较,siRNA组He La细胞的凋亡率增加,差异具有统计学意义(P<0.05);和阴性siRNA组及对照组比较,siRNA组HDAC1 mRNA表达量降低,差异具有统计学意义(P<0.05),siRNA+DDP组IC50值低于阴性siRNA+DDP组及对照+DDP组,差异具有统计学意义(P<0.05)。结论 HDAC1 siRNA可抑制HDAC1基因的表达水平,使组蛋白去乙酰化水平增加,提高宫颈癌He La细胞对顺铂的敏感性。
Objective To explore the application of ribonucleic acid（ RNA） in interfering the expression of histone acetylation enzyme 1（ HDAC1）,improving the sensitivity of human cervical carcinoma He La cells to cisplatin（ DDP）. Methods Human cervical cancer He La cells were transfected with liposome with small interfering RNA（ siRNA） of HDAC1. The cells were divided into the siRNA group,negative siRNA group and control group. The control,siRNA and negative siRNA groups were treated with DDP and assigned control ＋ DDP group,siRNA ＋ DDP and negative siRNA ＋ DDP groups respectively. Polymerase chain reaction（ PCR） was used to detect mRNA expression of HDAC1 gene,and flow cytometry was used to detect cell apoptosis rate,while methyl thiazolyl tetrazolium（ MTT） colorimetry was used to detect their sensitivity to DDP. Results Compared with negative siRNA group and control group,the apoptosis rate of He La cells in siRNA group increased,and the difference of them had statistical significance（ P〈0. 05）; HDAC1 expression of He La cells decreased,and the difference of them had statistical significance（ P〈0. 05）. Compared with negative siRNA ＋ DDP group and control ＋ DDP group,IC50 of siRNA ＋ DDP group reduced,and the difference of them had statistical significance（ P〈0. 05）. Conclusion HDAC1 siRNA can interfere and inhibit the expression of HDAC1 gene,increase the acetylation of histone,and improve the sensitivity of cervical cancer He La cells to cisplatin.
作者
方来福
钟国平
刘敏杰
解友邦
查桂平
叶小磊
FANG Lai-fu ZHONG Guo-ping LIU Min-jie XIE You-bang ZHA Gui-ping YE Xiao-lei(Department of Pathological, Yinzhou People＇ s Hospital, Ningbo, Zhejiang 315040, China)
出处
《中国卫生检验杂志》
CAS
2016年第22期3207-3209,3220,共4页
Chinese Journal of Health Laboratory Technology
基金
浙江省宁波市科技富民惠民项目(2015C50015)
浙江省宁波市自然科学基金项目(2013A610229)
浙江省宁波市自然科学基金项目(2016A610023)
作者简介
方来福（1979-），男，本科，主管技师，主要从事宫颈癌的基因及蛋白表达方面的研究。
通讯作者：叶小磊，E-mail：yexiaolei@nbu．edu．cn
