siRNA抑制HMGA1基因表达对LX-2细胞生物学功能的影响  

Effects of siRNA inhibit HMGA1 gene expression on LX-2 cell biological functions

作　　者：胡蕾[1] 刘莉[1] 张霜[1] 杜芳腾[1] 张吉翔[1] 

机构地区：[1]南昌大学第二附属医院消化内科/江西省分子医学重点实验室,南昌330006

出　　处：《重庆医学》2016年第17期2323-2326,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(81360074)

摘　　要：目的探讨高迁移率蛋白A1(HMGA1)siRNA对人肝星状细胞HMGA1、α-SMA、E-钙黏素(E-cadherin)基因的表达调控作用及增殖活性的影响,并探讨其可能的机制。方法将有效的人工合成的HMGA1siRNA转染人肝星状细胞LX-2(LX-2)后沉默HMGA1基因的表达。通过实时荧光定量PCR(RT-PCR)及蛋白免疫印迹法(Western blot)检测LX-2细胞中HMGA1、α-SMA和E-cadherin的mRNA及蛋白表达水平。采用四甲基偶氮唑盐(MTT)法检测LX-2细胞增殖水平。结果以HMGA1-siRNA序列1组沉默效果最好。TGF-β1刺激组与TGF-β1+NC-siRNA组之间细胞增殖水平、HMGA1、α-SMA、E-cadherin的基因及蛋白表达水平差异无统计学意义(P>0.05),细胞增殖水平、HMGA1、α-SMA表达均明显高于正常对照组(P<0.05),E-cadherin表达显著低于正常对照组(P<0.05);TGF-β1+HMGA1siRNA组细胞增殖水平及HMGA1、α-SMA的mRNA及蛋白表达水平较另外3组均显著下降(P<0.05),E-cadherin表达水平显著增高(P<0.05)。结论靶向HMGA1的siRNA能够沉默LX-2中HMGA1的表达;抑制TGF-β1诱导的LX-2增殖水平,提示HMGA1参与了TGF-β1诱导的肝星状细胞活化。Objective To investigate the effects of siRNA mediated HMGA 1 silence on proliferation and the gene expression of HMGA1 ,α-SMA and E-cadherin in activated hepatic stellate cells and its mechanisms .Methods Synthetic HMGA1 siRNA was transfected into LX-2 cells to silence the HMGA1 gene .The expression level of HMGA1 ,α-SMA and E-cadherin was determined by RT-PCR and Western blot experiments .LX-2 cell proliferation was assessed by M TT assay .Results The best inhibited effect was HMGA1-siRNA-1 .Compared with control group ,the cell proliferation and the mRNA and protein expression of HMGA 1 ,α-SMA in TGF-β1 group and TGF-β1 ＋ NC-siRNA group were significantly increased （P〈0 .05） ,without significant differences between the two groups （P〉0 .05） ,while the expression of E-cadherin in TGF-β1 group and TGF-β1 ＋ NC-siRNA group were significantly decreased compared with control group （P〈0 .05） .Meanwhile ,the cells in TGF-β1 ＋ HMGA1 siRNA group showed significantly decreased proliferation level ,down-regulated mRNA and protein expression of HMGA 1 ,α-SMA but up-regulated expression of E-cadherin compared with TGF-β1 group and TGF-β1 ＋ NC-siRNA group（P〈0 .05） .Conclusion HMGA1 interference could signifi-cantly down-regulate the expression of HMGA1 in LX-2 cells cultured with TGF-β1 ,thus inhibiting the proliferation and activation of the cells .

关 键 词：高迁移率蛋白A1 RNA干扰 肝星状细胞 肝纤维化 

分 类 号：R575.2[医药卫生—消化系统]