siRNA靶向沉默MALAT-1对鼻咽癌细胞增殖、凋亡及PI3K/ATK通路的影响  

Effects of siRNA targeting silencing MALAT-1 on proliferation,apoptosis and PI3K/ATK pathway of nasopharyngeal carcinoma cells

作　　者：曹亚莉[1] 李宏慧[1] 邵渊[1] 朱云[1] 黄建华 张鹏飞[1] Cao Yali;Li Honghui;Shao Yuan;Zhu Yun;Huang Jianhua;Zhang Pengfei(Department of Otolaryngology,Head and Neck Surgery,First Affiliated Hospital of Medical College of Xi'an Jiaotong University,Shaanxi Xi'an 710061,China.)

机构地区：[1]西安交通大学医学院第一附属医院耳鼻咽喉头颈外科

出　　处：《现代肿瘤医学》2020年第6期892-896,共5页Journal of Modern Oncology

基　　金：陕西省自然科学基础研究计划(面上项目)(编号:2017JM8072)

摘　　要：目的:探究siRNA靶向沉默MALAT-1对鼻咽癌细胞增殖、凋亡及PI3K/AKT通路的影响。方法:分别以10、20、30、40、50 nmol/L浓度的siRNA靶向沉默体外培养的人鼻咽癌细胞株CNE的基因MALAT-1,以实时荧光定量(qRT-PCR)分别在24、48 h后检测MALAT-1 mRNA表达水平,筛选合适的siRNA作用浓度和时间。将CNE细胞分为三组:空白对照组、siRNA阴性对照组和siRNA组,siRNA处理细胞后,采用CCK-8法检测细胞增殖情况,采用流式细胞技术检测细胞凋亡情况,Western blot检测各组细胞凋亡相关蛋白(Bcl-2、Bax、caspase-3)及PI3K/AKT通路蛋白(PI3K、ATK、p-AKT)表达情况。结果:选定30 nmol/L浓度的siRNA作用于CNE细胞48 h;siRNA处理细胞后,与空白对照组相比,siRNA组细胞增殖率明显降低,凋亡率明显升高,Bax、caspase-3蛋白表达明显升高,Bcl-2、PI3K、p-AKT蛋白表达明显降低,差异均有统计学意义(P<0.05);AKT蛋白表达无明显变化,差异无统计学意义(P>0.05)。siRNA阴性对照组各指标均无明显变化,差异无统计学意义(P>0.05)。结论:siRNA可靶向沉默MALAT-1,抑制鼻咽癌细胞增殖,促进其凋亡,可能是通过抑制PI3K/AKT通路实现的。Objective:To investigate the effects of siRNA targeting silencing MALAT-1 on proliferation,apoptosis and PI3K/AKT pathway of nasopharyngeal carcinoma cells.Methods:The MALAT-1 gene of human nasopharyngeal carcinoma cell line CNE was targeted silenced with siRNA at concentrations of 10,20,30,40 and 50 nmol/L,respectively.Real-time fluorescence quantitative analysis(qRT-PCR)was used to detect the expression of MALAT-1 after 24 hours and 48 hours respectively,screened suitable concentration and time of siRNA treatment.CNE cells were divided into three groups:Blank control group,siRNA negative control group and siRNA group.After siRNA treatment of cells,CCK-8 method was used to detect cell proliferation.Cell apoptosis was detected by flow cytometry.Western blot was used to detect the expressions of apoptosis-related proteins(Bcl-2,Bax,caspase-3),PI3K/AKT pathway proteins(PI3K,ATK,p-AKT).Results:30 nmol/L siRNA was selected to act on CNE cells for 48 hours.Compared with the blank control group,the proliferation rate of cells treated with siRNA decreased significantly.The apoptotic rate and the expressions of Bax and caspase-3 proteins were significantly increased.The expressions of Bcl-2,PI3K and p-AKT proteins were significantly decreased(P<0.05).There was no significant change in AKT protein expression(P>0.05).There was no significant difference in the indexes of siRNA negative control group(P>0.05).Conclusion:siRNA can silence MALAT-1,inhibit the proliferation and promote apoptosis of nasopharyngeal carcinoma cells,probably by inhibiting PI3K/AKT pathway.

关 键 词：SIRNA沉默 增殖 凋亡 PI3K/AKT通路 MALAT-1 鼻咽癌细胞 

分 类 号：R739.6[医药卫生—肿瘤]