肝再生增强因子RNAi对肝癌细胞株HepG2表达MHCI、DR、CD80、CD86的影响  

Effects of Human Augmenter of Liver Regeneration RNA Interference on Expression of MHCI,DR,CD80,CD86 in HepG2 Cells

作　　者：谢松丽[1] 张云霞[1] 孙航[1] 刘杞[1] 

机构地区：[1]重庆医科大学附属第二医院肝病研究所,重庆400010

出　　处：《生物技术通报》2009年第6期122-126,共5页Biotechnology Bulletin

基　　金：国家自然科学基金(30570826)

摘　　要：利用肝再生增强因子(augmenter of liver regeneration,ALR)的siRNA阻断人肝癌细胞株HepG2表达ALR,观察HepG2细胞MHCI、DR、CD80、CD86的表达是否受影响,以探讨ALR是否通过影响MHCI、DR、CD80、CD86表达参与肝癌细胞逃避机体的免疫监视。培养HepG2,分为未转染组、转染阳性SiRNA组、转染阴性SiRNA组、转染脂质体空质粒组;RT-PCR检测转染前后hALR mRNA的表达,RT-PCR、Western blot印迹法、流式细胞术(FCM)检测HepG2细胞表达MHCI、DR、CD80、CD86的水平。结果表明,转染阳性SiRNA组hALR mRNA的表达较转染阴性SiRNA组明显下降,转染阳性SiRNA组和转染阴性SiRNA组MHCI、DR、CD80、CD86的表达均无差异;脂质体转染组除CD80 mRNA外,MHCI、DR、CD86 mRNA表达均较未转染组增高;脂质体转染组MHCI、DR、CD80、CD86的表达水平均较未转染组有上升趋势。因此,阳性SiRNA具有显著和特异性抑制hALR表达的作用,阻断ALR的表达不影响HepG2细胞表达MHCI、DR、CD80、CD86,ALR不通过影响MHCI、DR、CD80、CD86的表达参与肝癌细胞逃避机体免疫监视。转染载体可能影响某些基因的表达。This study was to investigate the effect of hALR on the expression of MHCI, DR, CD80 and CD86 in HepG2 cells, by which ALR could participate HCC escape from immunosurveillance HepG2 cells are divided, into four groups, including those transfected with RNAi plasmid pSIALR-A, transfected with control plasmid pSIALR-B, transfected with lipofectamine and untransfected. RT- PCR, FCM and Western blot were involved in to detect MHCI, DR, CD80 and CD86. Results indicated that hALR mRNA in pSIALRA group was lower than pSIALR-B group. The differences between MHCI, DR, CD80, and CD86 in pSIALR-A group and pSIALR-B group were not significant. In lipofectamin group, CD80 mRNA was lower and MHCI, DR and CD86 mRNA were higher than untransfected group using RT-PCR. MHCI, DR, CD80 and CD86 were higher than untransfected group using FCM. Thus, pSIALR-A can inhibit the expression of ALR significantly and specifically. Blocking the expression of hALR by siRNA could not affect the expression of MHCI, DR, CD80 and CD86 in HepG2 cells. ALR fail to participate HCC immune escape by affect the expression of MHCI, DR, CD80, CD86. Transfection carrier might affect expression of some genes.

关 键 词：人肝再生增强因子 RNA干扰 主要组织相容性复合体B7 

分 类 号：R735.7[医药卫生—肿瘤]