Supplementary Material for: LncRNA HANR Promotes Tumorigenesis and Increase of Chemoresistance in Hepatocellular Carcinoma

<p><b><i>Background/Aims:</i></b> Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world and the third leading cause of cancer-related death. Critical roles for long non-coding RNAs (lncRNAs) have recently been demonstrated for a variety of cancers, including hepatocellular carcinoma. However, the effect and mechanism of lncRNAs in HCC tumorigenesis and chemoresistance have not been extensively characterized. <b><i>Methods:</i></b> In the current study, we have identified a HCC-expressed lncRNA termed as <i>HANR</i> (HCC associated long non-coding RNA). We identified <i>HANR</i> by microarray analysis and validated its up-regulated expression by quantitative PCR. RNA pull-down and pathway analyses were conducted to evaluate physical and functional interactions with <i>HANR</i>. <i>In vivo</i> experiments were performed to assess tumorigenesis and increase of chemoresistance. In addition, the <i>HANR</i> expression in HCC specimens was detected by FISH. Xenograft and orthotopic mice model was constructed to observe the effect of <i>HANR</i> on tumorigenesis and chemoresistance <i>in vivo</i>. <b><i>Results:</i></b> <i>HANR</i> was demonstrated to be up-regulated in HCC patients and HCC cell lines. Increased <i>HANR</i> expression in HCC predicted short survival of patients. Knock-down of <i>HANR</i> markedly retarded cell proliferation, suppressed HCC xenograft/orthotopic tumor growth, induced apoptosis and enhanced chemosensitivity to doxorubicin, while over-expression of <i>HANR</i> showed the opposite effects. It was found that <i>HANR</i> bind to GSKIP for regulating the phosphorylation of GSK3β in HCC. <b><i>Conclusion:</i></b> Our results demonstrate that <i>HANR</i> contributes to the development of HCC and is a promising therapeutic target for chemosensitization of HCC cells to doxorubicin, which may represent a promising therapeutic target in the future.</p>