摘要
目的二甲双胍可抑制鼻咽癌细胞的增殖,但作用机制尚不清楚。文章研究二甲双胍对鼻咽癌细胞CNE-1增殖与凋亡的效应,并探讨miR-let-7a、IGF-1R在其中的作用。方法分别以不同浓度二甲双胍(0、5、10、20、40、80mmol/L)干预鼻咽癌细胞CNE-124h后,采用CCK8法检测细胞增殖活性;流式细胞仪技术分析细胞凋亡率;实时荧光定量PCR(qPCR)检测bcl-2、bax、IGF-1RmRNA表达水平,miR-let-7a的表达水平;Westernblot方法检测IGF-1R蛋白的表达。结果CCK8结果显示,各浓度组二甲双胍处理细胞24h后,二甲双胍可抑制CNE-1细胞的增殖活性,随药物浓度增高抑制作用呈增强趋势,差异均有统计学意义(P<0.05)。流式细胞仪凋亡分析结果显示,0、5、10、20、40mmol/L二甲双胍处理后总凋亡率分别为(6.17±0.74)%、(8.11±1.29)%、(11.97±3.82)%、(13.94±2.98)%、(24.2±2.79)%;与0mmol/L二甲双胍处理比较相比,10、20、40mmol/L二甲双胍处理后凋亡率随浓度升高而呈上升趋势(P<0.05)。qPCR结果显示,0、5、10、20mmol/L浓度二甲双胍作用后24h后,CNE-1细胞的bcl-2、bcl-2/bax、IGF-1R、miR-let-7amRNA表达水平随二甲双胍浓度增加而逐渐下调,而bax基因表达水平则上调,与0mmol/L二甲双胍作用后相比,差异均有统计学意义(P<0.05)。结论二甲双胍有抑制鼻咽癌细胞CNE-1增殖并诱导其凋亡的作用,其机制可能与miR-let-7a表达上调、IGF-1R表达下调有关。
Objective research the effects of metformin on proliferation and apoptosis in nasopharyngeal carcinoma cell CNE-1 and investigate the role of miR-let-7a、IGF-1R in it. Methods The nasopharyngeal carcinoma cell CNE-1 was treated with different concentrations of metformin for 24h, then the proliferation activity of cell was detected by CCK8 method;the apoptosis rate of cell was measured by flow cytometry;the expression levels of bcl-2、bax and IGF-1R mRNA and miR-let-7a were detected by real-time quantitative PCR;the expression level of IGF-1R protein was detected by Western blot. Results Compared with the control group, the cell proliferation activity of metformin group decreased(P<0.05), and it gradually decreased along with the increase of metformin concentration. The cell apoptosis rate of metformin group increased(P<0.05 except for the 5 mmol/L group), and it gradually increased along with the increase of metformin concentration. The expression levels of bax mRNA and miR-let-7a were up-regulated in the metformin group(P<0.05), while the expression levels of bcl-2 mRNA,IGF-1R mRNA and IGF-1R protein were decreased(P<0.05). Conclusion Metformin could inhibit the proliferation and induce apoptosis of CNE-1. The mechanism maybe related to the up-regulation of miR-let-7a and down-regulation of IGF-1R.
作者
汤燕琴
胡兵
余杰情
范立
樊欣
陈剑飞
张志远
TANG Yan-qin;HU Bing;YU Jie-qing;FAN Li;FAN Xin;CHEN Jian-fei;ZHANG Zhi-yuan(2.Medical School,Shenzhen University,Shenzhen 518000,Guangdong,China)
出处
《医学研究生学报》
CAS
北大核心
2019年第9期926-930,共5页
Journal of Medical Postgraduates
基金
江西省科技厅重点研发项目(20171BBG70006)
