伊立替康调控SNHG1/miR-195的表达抑制肝癌细胞增殖、迁移、侵袭的影响及机制研究  

Effects of irinotecan on the proliferation,migration and invasion of hepatoma cells by regulating the expressions of SNHG1/miR195 and its action mechanism in vitro

作　　者：孙绪举[1] 干飞[2] 胡娟[3] 张继东[2] SUN Xuju;GAN Fei;HU Juan(Department of Pharmacy,Liyuan Hospital Affiliated to Tongji Medical College of Huazhong Unicersity of Science and Technology,Hubei,Wuhan 430077,China;不详)

机构地区：[1]华中科技大学同济医学院附属梨园医院药剂科,武汉市430077 [2]华中科技大学医院药剂科,430074 [3]华中科技大学医院外科

出　　处：《河北医药》2020年第24期3691-3696,共6页Hebei Medical Journal

摘　　要：目的研究伊立替康对肝癌细胞增殖、迁移、侵袭的影响及其作用机制。方法以16 ml/L、32 ml/L、64 ml/L浓度的伊立替康处理肝癌细胞HepG2,MTT法检测细胞增殖,Transwell小室法检测细胞迁移和侵袭,Western blot检测细胞周期蛋白D1(CyclinD1)、p21、E-钙粘蛋白(E-cadherin)、基质金属蛋白酶-2(MMP-2)表达,qPCR检测SNHG1和miR-195表达。构建抑制SNHG1表达的HepG2细胞,或构建过表达SNHG1的细胞并进行32 ml/L伊立替康处理,观察其对细胞增殖、迁移和侵袭的影响。StarBase软件预测出miR-195是SNHG1的靶基因,双荧光素酶报告实验进一步验证。细胞HepG2中转染anti-miR-195并使用32 ml/L伊立替康处理,检测细胞增殖、迁移和侵袭。结果不同浓度伊立替康明显降低HepG2细胞活性,并呈剂量和时间依赖性。32 ml/L伊立替康显著减少迁移细胞数、侵袭细胞数、Cyclin D1蛋白、MMP-2蛋白、SNHG1表达量,明显提高P21、E-cadherin蛋白水平(P<0.05)。抑制SNHG1表达显著降低24 h、48 h和72 h的细胞活性,减少迁移细胞数侵袭细胞数、Cyclin D1、MMP-2蛋白表达量,提升P21、E-cadherin蛋白水平(P<0.05)。过表达SNHG1逆转了伊立替康对HepG2细胞增殖、迁移、侵袭、Cyclin D1、MMP-2蛋白表达的抑制作用,以及逆转对P21、E-cadherin蛋白表达的促进作用。SNHG1靶向调控miR-195表达。抑制miR-195表达逆转伊立替康抑制HepG2细胞增殖、迁移、侵袭的作用。结论伊立替康通过调控SNHG1/miR-195表达抑制肝癌细胞增殖、迁移、侵袭。Objective To investigate the effects of irinotecan on the proliferation,migration and invasion of hepatoma cells by regulating the expressions of SNHG1/miR195 and its action mechanism in vitro.Methods HepG2 cells were treated by irinotecan at the concentration of 16ml/L,32ml/L and 64ml/L in vitro.The cell proliferation was detected by MTT assay,cell migration and invasion were determined by Transwell chamber assay.The protein expressions of CyclinD1,p21,Ecadherin and matrix metalloproteinase 2(MMP-2)were detected by Western Blot.The expression levels of SNHG1 and miR195 were detected by qPCR.Moreover the HepG2 cells inhibiting SNHG1 expression or overexpression SNHG1 were established,which were treated by 32ml/L irinotecan,and its effects on cell’s proliferation,migration and invasion were observed.The starBase software predicted that miR195 was the target gene of SNHG1,and the dual luciferase reporter assay was used to further validate the results.The HepG2 cells were transfected with anti-miR195 and treated by 32ml/L irinotecan,then,the cell’s proliferation,migration and invasion were detected.Results The different concentrations of irinotecan significantly reduced the activity of HepG2 cells in a dose and time dependent manner.The 32ml/L irinotecan obviously reduced the number of migrated cells,the number of invasive cells,the protein expressions of Cyclin D1,MMP2 and SNHG1,which significantly increased the protein levels of P21 and Ecadherin(P<0.05).The inhibition of SNHG1 expression significantly decreased cell viability at 24h,48h and 72h after treatment,reduced the number of invasive cells,and the protein expressions of Cyclin D1 and MMP2,however,which increased the protein levels of P21 and Ecadherin(P<0.05).The overexpression of SNHG1 reversed the inhibitory effects of irinotecan on cell proliferation,migration,invasion,and on the protein expressions of Cyclin D1 and MMP2 in HepG2 cells,which reversed the promotion effects on the protein expressions of P21 and Ecadherin.In addition SNHG1 could target

关 键 词：伊立替康 SNHG1 miR-195 肝癌 增殖 

分 类 号：R735.7[医药卫生—肿瘤]