基于STAT3通路探讨沉默星形胶质上调基因-1对人胆囊癌GBC-SD细胞增殖及凋亡的影响  

Discussion on the effect of ailencing astrocyte elevated gene-1 on the proliferation and apoptosis of human gallbladder cancer GBC-SD cells based on the STAT3 pathway

作　　者：徐松涛[1] 孟松[1] 顾凤华[2] 顾国建[2] XU Songtao;MENG Song;GU Fenghua;GU Guojian(Luohe Medical College,Luohe 462002,Henan,China;Taicang Hospital Affiliated to Suzhou University,Taicang 215400,Jiangsu,China)

机构地区：[1]漯河医学高等专科学校,河南漯河462002 [2]苏州大学附属太仓医院,江苏太仓215400

出　　处：《现代中西医结合杂志》2020年第27期2970-2974,共5页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：江苏省太仓市基础研究计划项目(TC2018JCYL19);漯河医学高等专科学校创新创业发展能力提升工程项目(2019-LYZKYZD018)。

摘　　要：目的探讨沉默星形胶质细胞上调基因-1(AEG-1)对人胆囊癌GBC-SD细胞增殖、凋亡及STAT3信号通路的影响。方法实验分为空白对照组(仅有空质粒,无任何阴性或阳性干扰序列)、阴性对照组(携带AEG-1阴性序列)、RNA干扰组(携带AEG-1基因干扰序列),将人GBC-SD细胞接种于培养板内,按要求进行质粒载体转染细胞操作,采用qPCR法检测各组AEG-1 mRNA表达水平,采用Western blot法检测STAT3、p-STAT3和AEG-1蛋白表达水平,采用MTT法检测各组GBC-SD细胞增殖情况,采用Hoechst 33342染色法检测各组GBC-SD细胞凋亡情况,采用ELISA法检测各组GBC-SD细胞分泌基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)水平。结果与空白对照组和阴性对照组比较,RNA干扰组AEG-1 mRNA及蛋白表达水平和p-STAT3蛋白表达水平均明显降低,GBC-SD细胞增殖明显下降,凋亡率明显增高,MMP-2和MMP-9水平明显降低,差异均有统计学意义(P均<0.05)。结论沉默AEG-1可以通过STAT3信号通路抑制人GBC-SD细胞的增殖和诱导其凋亡。Objective It is to investigate the effects of silencing astrocyte elevated gene-1(AEG-1)on the proliferation and apoptosis of human gallbladder cancer GBC-SD cells based on the STAT3 pathway.Methods The experiment was divided into blank control group(only had empty plasmid,without any negative or positive interference sequence),negative control group(with AEG-1 negative sequence),RNA interference group(with AEG-1 gene interference sequence),the human GBC-SD cells were inoculated in a culture plate,and plasmid vector transfection was performed as required.The expression levels of AEG-1 mRNA in each group were detected by qPCR method,and the protein expression levels of STAT3,p-STAT3 and AEG-1 were detected by Western blot,the proliferation of GBC-SD cells in each group was detected by MTT method,the apoptosis of GBC-SD cells in each group was detected by Hoechst 33342 staining method,and the levels of matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-2)secreted by the GBC-SD cells were detected by ELISA method.Results Compared with the blank control group and the negative control group,the levels of AEG-1 mRNA and protein expression and p-STAT3 protein expression in the RNA interference group were significantly reduced,the proliferation of GBC-SD cells was significantly decreased,and the apoptosis rate was significantly increased,the levels of MMP-2 and MMP-9 were significantly reduced,and the differences were statistically significant(P<0.05).Conclusion Silencing AEG-1 can inhibit the proliferation and induce apoptosis of human GBC-SD cells through the STAT3 signaling pathway.

关 键 词：胆囊癌 星形胶质细胞上调基因-1 质粒 基质金属蛋白酶 

分 类 号：R735.8[医药卫生—肿瘤]