浙江医学2017，Vol.39Issue(4)：250-254,5.DOI:10.12056/j.issn.1006-2785.2017.39.4.2016-953
长期索拉菲尼暴露促进Huh7肝癌细胞上皮间质变的实验研究
Epithelial-mesenchymal transition and migrant/invasion ability in human sorafenib-resistant hepatocellular carcinoma Huh7 cells
摘要
Abstract
Objective To investigate epithelial-mesenchymal transition (EMT) and migrant/invasion ability in sorafenibresistant human hepatocellular carcinoma cells (Huh7R).Methods The Huh7R was induced by long-term exposure to low dose sorafenib.The morphological changes of Huh7R was observed with phase contrast microscopy,the drug sensitivity was evaluated with cytotoxicity cell counting kit-8 (CCK-8) assay;the expression of ABC family resistance genes ABCB1,ABCC1,4BCG2,and transcription factor genes Snail,Slug was detected with real-time PCR (RT-PCR),the resistance related protein ABCG2,EMT marker E-cadherin,Vimentin and N-cadherin,transcription factors Snail and Slug was detected with Western blot.Migration and invasion properties of Huh7R cells were assessed using Transwell assays.Results Phase contrast microscopy showed that the Huh7R cells were elongated with fibrous projections and the morphological changes were consistent with typical EMT changes.CCK-8 results showed that the Huh7R cells were less sensitive to sorafenib than parental Huh7 cells.Fluorescence quantitative PCR results showed that compared to the parental Huh7 cells,the expression of resistance gene ABCB1,ABCC1 and transcription factor Snail,Slug in Huh7R cells was up-regulated significantly,and Western blot results showed that the expression of ABCG2 protein was significantly increased,the expression of epithelial marker E-cadherin protein was decreased,and the expression of mesenchymal marker protein Vimentin and N-cadherin increased,the expression levels of snail and slug were up-regulated.Transwell invasion assay indicated that Huh7R migration and invasion ability was increased compared with parental cells.Conclusion Theresultsindicatethatlongtermexposuretolowdosesorafenibpromotestheexpression of snail and Slug in human heparocellular carcinoma Huh7R cells,which mediates EMT and enhances the migrant and invasion ability of Huh7R cells.关键词
肝癌/索拉菲尼/转录因子/上皮间质变/耐药Key words
Hepatocellular carcinoma/Sorafenib/Transcription factor/Epithelial mesenchymal transition/Drug resistance引用本文复制引用
刘俊,王清清,曹浩强,张浩..长期索拉菲尼暴露促进Huh7肝癌细胞上皮间质变的实验研究[J].浙江医学,2017,39(4):250-254,5.基金项目
浙江省“十二五”基层卫生适宜技术成果转化工程重大项目(2013T301-12、15) （2013T301-12、15）
嘉兴市科技计划项目(2013AY21042-5) （2013AY21042-5）
嘉兴市重点科技创新团队项目([2013]3号) （[2013]3号）
