LncRNA regulated by hnRNPAB in HCC cell lines. LncRNA regulated by hnRNPAB in HCC cell lines

To reveal the HNRNPAB-regulated lncRNAs, we performed microarray analyses to screen differential lncRNAs in HCC cells after stable overexpression or knockdown of HNRNPAB. MHCC97H and HCCLM3 (HCC cell lines with high-metastatic potentials), PLC/PRF/5 and HepG2 (HCC cell lines with low-metastatic potentials) were used in this study. HepG2-control, HepG2-hnRNPAB, PLC/PRF/5-control, PLC/PRF/5-hnRNPAB, MHCC97H-control, MHCC97H-sh-hnRNPAB, HCCLM3-control, HCCLM3-sh-hnRNPAB were perpared with hU6-MCS-CBh-gcGFP-IRES-puromycin-shRNA-HNRNPAB/mock lentiviral and Ubi-MCS-SV40-EGFP-IRES-puromycin-HNRNPAB/mock cDNA lentiviral,respectively, each performed in triplicate. Overall design: HepG2-control, HepG2-hnRNPAB, PLC/PRF/5-control, PLC/PRF/5-hnRNPAB, MHCC97H-control, MHCC97H-sh-hnRNPAB, HCCLM3-control, HCCLM3-sh-hnRNPAB were perpared and validated by qRT-PCR. Independent triplicate RNA samples in each cell was subjected to lncRNA microarrays.

为揭示受异质核糖核蛋白AB（HNRNPAB）调控的长链非编码RNA（lncRNAs），本研究通过长链非编码RNA基因芯片分析，筛选稳定过表达或敲低HNRNPAB的肝细胞癌（HCC）细胞中差异表达的lncRNAs。本研究选用高转移潜能HCC细胞系MHCC97H与HCCLM3，以及低转移潜能HCC细胞系PLC/PRF/5与HepG2。分别采用携带Ubi-MCS-SV40-EGFP-IRES-puromycin-HNRNPAB/mock cDNA的慢病毒构建HepG2对照组、HepG2-hnRNPAB过表达组、PLC/PRF/5对照组、PLC/PRF/5-hnRNPAB过表达组；采用携带hU6-MCS-CBh-gcGFP-IRES-puromycin-shRNA-HNRNPAB/mock的慢病毒构建MHCC97H对照组、MHCC97H-sh-hnRNPAB敲低组、HCCLM3对照组、HCCLM3-sh-hnRNPAB敲低组，每组均设置3次生物学重复。总体实验设计如下：上述各组细胞均已完成构建并经实时荧光定量PCR（qRT-PCR）验证，随后提取每组细胞的独立三份RNA重复样本，用于长链非编码RNA基因芯片检测。