Abstract
Long non-coding RNAs (lncRNAs) play key roles in cancer initiation and progression. The aim was to investigate the biological functions and clinical significance of long non-coding RNA CARLo-5 in hepatocellular carcinoma (HCC). QRT-PCR was performed to investigate CARLo-5 expression in HCC tissues and cells. Kaplan-Meier curve and multivariate analysis validated the association between CARLo-5 expression and overall survival (OS) in HCC patients. Cell proliferation and invasion was performed by CCK8 cell proliferation, cell colony formation and transwell invasion assays. Western-blot assay was performed to evaluate the protein expression of Twist1, ZEB1, E-cadherin and Vimentin. Tumor xenografts were performed to evaluate the effect of CARLo-5 on tumor growth in vivo. RNA Immunoprecipitation (RIP) and Chromatin Immunoprecipitation (ChIP) were also performed. Our results showed that CARLo-5 expression was significantly higher in HCC tissues and upregulated CARLo-5 expression was closely correlated with tumor size and advanced tumor stage. Kaplan-Meier curve and multivariate analysis validated that higher CARLo-5 expression predicted a poor prognosis for HCC patients and was an independent risk factor for OS in HCC patients. In vitro, knockdown of CARLo-5 inhibited cell proliferation, colony formation, cell invasion and inhibited the cell epithelial-mesenchymal transition (EMT) by up-regulating the E-cadherin expression and down-regulating Twist1, ZEB1 and vimentin expression in HCC cells. Furthermore, we demonstrated that CARLo-5 inhibited the miR-200b expression via EZH2. In vivo, knockdown of CARLo-5 significantly inhibited the tumor growth. Thus, our results indicated that CARLo-5 represented a novel tumor biomarker and therapeutic target for HCC.
Highlights
Hepatocellular carcinoma (HCC) is the most commonly diagnosed malignancy in the liver and ranks the third most frequent cancer-related mortality in the world [1, 2]
According the median expression of Cancer-associated region long noncoding RNAs (CARLos)-5 in hepatocellular carcinoma (HCC) tissues (2.45), the CARLo-5 expression levels were classified two groups, the results showed that CARLo-5 expression was positively correlated with the larger tumor size and American Joint Committee on Cancer (AJCC) stage, but no correlation with age, gender, and so on (P
The results demonstrated that higher expression of CARLo-5 in HCC patients predicted a poor prognosis for HCC patients (Figure 1B)
Summary
Hepatocellular carcinoma (HCC) is the most commonly diagnosed malignancy in the liver and ranks the third most frequent cancer-related mortality in the world [1, 2]. Abnormal expression www.impactjournals.com/oncotarget of lncRNAs have been found to be associated with hepatocarcinogenesis and play key roles in progression and metastasis [6]. Up-regulated in liver cancer (HULC) was originally identified as the most overexpressed long non-coding RNA in hepatocellular carcinoma. Patients with HOTAIR expression had significantly poorer over survival time and a larger primary tumor size than those without HOTAIR expression [9]. These above results indicated LncRNAs play potential roles in HCC
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