miR-23a/SATB2信号通路对结直肠癌SW480细胞凋亡和转移的影响  

Effect of miR-23a/SATB2 signaling pathway on apoptosis and metastasis of colorectal cancer SW480 cells

作　　者：高永涛[1] 白浪[1] 樊华[1] 刘涛[1] 姬乐[1] 雷星[1] 邓世波 袁江涛[1] GAO Yongtao;BAI Lang;FAN Hua;LIU Tao;JI Le;LEI Xing;DENG Shibo;YUAN Jiangtao(Department of Gastrointestinal Surgery,Yan′an University Affiliated Hospital,Yan′an 716000,China)

机构地区：[1]延安大学附属医院胃肠外科,716000

出　　处：《国际消化病杂志》2021年第4期289-295,共7页International Journal of Digestive Diseases

基　　金：延安市科学技术研究发展计划项目(2013-kw06)。

摘　　要：目的探讨miR-23a/SATB2信号通路对结直肠癌SW480细胞凋亡和转移的影响。方法在结直肠癌SW480细胞中转染miR-23a抑制剂(inhibitor),采用定量RT-PCR(qRT-PCR)法检测miR-23a mRNA相对表达量,CCK-8法检测细胞增殖,平板克隆法检测细胞克隆能力,流式细胞术测定细胞凋亡,Transwell小室检测细胞侵袭和迁移,蛋白质印迹法(Western blotting)检测基质金属蛋白酶-2(MMP-2)、剪切的天冬氨酸特异性半胱氨酸蛋白酶-3(C-Caspase-3)蛋白表达水平。靶基因预测软件预测SATB2可能成为miR-23a的靶基因,荧光素酶报告系统鉴定靶向关系。在细胞中共转染SATB2小干扰RNA(siRNA)和miR-23a inhibitor,采用上述方法检测细胞增殖、克隆、凋亡、侵袭、迁移和MMP-2、C-Caspase-3蛋白表达水平的变化。结果转染miR-23a inhibitor可明显降低结直肠癌SW480细胞中miR-23a mRNA的相对表达量,抑制细胞增殖、克隆、侵袭、迁移和MMP-2蛋白表达,促进细胞凋亡和C-Caspase-3蛋白表达。SATB2受到miR-23a的靶向负调控作用。SATB2 siRNA可逆转下调miR-23a表达对结直肠癌SW480细胞增殖、克隆、侵袭、迁移的抑制作用和凋亡诱导作用。结论miR-23a/SATB2信号通路具有调控结直肠癌SW480细胞凋亡和转移的潜能。Objective This paper intends to explore the effect of miR-23a/SATB2 signaling pathway on apoptosis and metastasis of colorectal cancer SW480 cells.Methods The miR-23a inhibitor was transfected into colorectal cancer SW480 cells,the expression of miR-23a mRNA was detected by qRT-PCR,the cell proliferation was detected by CCK-8,the cell cloning ability was detected by plate cloning,the cell apoptosis was measured by flow cytometry,the cell invasion and migration were detected by transwell chamber,and the expressions of MMP-2 and C-Caspase-3 were detected by western blotting.Target gene prediction software predicts that SATB2 may be a target gene of miR-23a.The targeting relationship was identified by the luciferase reporting system.The co-transfection of SATB2 siRNA and miR-23a inhibitor in cells,cell proliferation,cloning,apoptosis,invasion,migration and expression of MMP-2 and C-caspase-3 were detected by using the above methods.Results The transfection of miR-23a inhibitor is found to be able to significantly reduce the expression of miR-23a mRNA in colorectal cancer SW480 cells,inhibiting cell proliferation,cloning,invasion,migration and expression of MMP-2 protein,and promoting apoptosis and C-Caspase-3 protein expression.SATB2 is negatively regulated by the targeting of miR-23a.SATB2 siRNA can reverse the inhibitory effect of down-regulation of miR-23a on proliferation,cloning,invasion,migration and apoptosis induction of colorectal cancer SW480 cells.Conclusions The miR-23a/SATB2 signaling pathway has the potential to regulate apoptosis and metastasis of colorectal cancer SW480 cells.

关 键 词：miR-23a SATB2 结直肠癌 凋亡 

分 类 号：R735.34[医药卫生—肿瘤]