摘要
目的探讨差异表达的长链非编码RNA(lncRNA)对肝癌索拉菲尼耐药细胞迁移、侵袭的影响。方法采用lncRNA芯片微阵列分析肝癌索拉菲尼耐药细胞Huh7SR和亲本细胞Huh7中的lncRNA,筛选出差异表达lncRNA;荧光定量PCR法进一步验证差异表达的lncRNA。将转染后的肝癌索拉菲尼耐药细胞Huh7SR分为ENST00000484679-小干扰RNA(siRNA)组和阴性对照(siNC)组。ENST00000484679-siRNA组加入lncRNA ENST00000484679-siRNA转染,siNC组加入无关序列NC-siRNA转染。Transwell实验观察肝癌索拉菲尼耐药细胞株迁移侵袭能力的变化;Western blot法检测肝癌索拉菲尼耐药细胞株的上皮-间质转化(EMT)标记蛋白钙黏蛋白N(N-cadherin)、钙黏蛋白E(E-cadherin)、波形蛋白(Vimentin)表达水平。结果芯片筛选出lncRNA ENST00000484679在肝癌索拉菲尼耐药细胞株中高表达,荧光定量PCR进一步验证了Huh7SR细胞ENST00000484679 mRNA表达水平高于Huh7细胞(P<0.01)。ENST00000484679-siRNA组ENST00000484679 mRNA表达水平低于siNC组,迁移细胞数和侵袭细胞数均低于siNC组,N-cadherin、Vimentin蛋白表达水平均低于siNC组,E-cadherin蛋白表达水平高于siNC组,差异均有统计学意义(均P<0.05)。结论lncRNA ENST00000484679促进肝癌索拉菲尼耐药细胞的迁移和侵袭能力可能与EMT有关。
Objective To investigate the effect of differentially expressed lncRNA on the migration and invasion of sorafenib-resistant hepatocellular carcinoma cells.Methods lncRNA microarray was used to analyze lncRNA in sorafenib resistant hepatocellular carcinoma cells Huh7SR and parental cells Huh7,and to screen out differentially expressed lncRNA.The differentially expressed lncRNA was further verified by fluorescence quantitative PCR.The transfected sorafenib-resistant hepatocellular carcinoma cells Huh7SR were divided into ENST00000484679-siRNA group and siNC group.ENST 00000484679-siRNA group was transfected with lncRNA ENST00000484679-siRNA,and siNC group was transfected with unrelated sequence NC-siRNA.Transwell assay was used to observe the changes of migration and invasion ability of sorafenib-resistant hepatocellular carcinoma cell lines.Western blot was used to detect the expression levels of epithelial mesenchymal transition(EMT)marker proteins N-cadherin,E-cadherin and Vimentin in sorafenib-resistant cell lines.Results The microarray showed that lncRNA ENST00000484679 was highly expressed in Huh7SR cells,which was further verified by fluorescence quantitative PCR(P<0.01).The mRNA expression level of ENST00000484679 in the ENST00000484679-siRNA group was lower than that in the siNC group,the number of migrating cells and invading cells were lower than that in the siNC group,and the protein expression levels of N-cadherin and Vimentin were lower than that in the siNC group,the expression level of E-cadherin protein was higher than that in the siNC group(all P<0.05).Conclusion lncRNA ENST00000484679 promotes the migration and invasion of sorafenib-resistant hepatocellular carcinoma cells,which may be related to EMT.
作者
王清清
张杰
WANG Qingqing;ZHANG Jie(Department of Hepatobiliary and Pancreatic Surgery,the First Hospital of Jiaxing,Jiaxing 314001,China;不详)
出处
《浙江医学》
CAS
2022年第20期2144-2148,共5页
Zhejiang Medical Journal
基金
浙江省卫生健康科技计划项目(2021KY1101)
嘉兴市科技计划项目(2019AY32029)。
关键词
长链非编码RNA
肝癌
索拉菲尼耐药
上皮-间质转化
Long non-coding RNA
Liver cancer
Sorafenib-resistance
Epithelial-mesenchymal transition
