LncRNA HULC shRNA disinhibits miR-377-5p to suppress the growth and invasion of hepatocellular carcinoma in vitro and hepatocarcinogenesis in vivo

Abstract

Background: Aberrant expression of up-regulated long non-coding RNA [LncRNA highly upregulated in liver cancer (HULC)] has been observed to play an important regulatory role in the development of multiple human diseases. However, the molecular mechanism underlying the role of HULC and miR-377-5p in HCC needs to be urgently explored. Methods:The mRNA and protein expression levels of HULC were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot in hepatocellular carcinoma (HCC) cell line HB611, HepG2 and H22, respectively. HULC-shRNA was transfected into HepG-2 cells, which were randomly divided into the control, shRNA-NC, and sh-HULC groups. The correlation between HULC and miR-377-5p was analyzed by performing a luciferase reporter assay. The targeting relationship between miR-377-5p and hypoxia-inhibitory factor-1α (HIF-1α) was also investigated using a luciferase reporter assay.Sh-HULC and miR-377-5p inhibitors were transfected either alone or together into HepG2 cells, and which were divided into the control group, the sh-HULC group, the miR-377-5p inhibitor, and the sh-HULC + inhibitor group for subsequent experiments. HepG2 cell proliferation and invasion were measured by 5-Ethynyl-2-Deoxyuridine (EdU) staining and Transwell invasion assay, respectively. Western plot was carried out to detect the protein expression levels of Ki67, PCNA, E-cadherin, and N-cadherin.Tumor xenograft mouse models were established to confirm the effect of HULC down-regulation on the development of HCC in vivo.Results: The mRNA and protein expression levels of HULC were markedly increased, whereas the mRNA expression levels of miR-377-5p were decreased in HCC cell lines. HepG2 cell proliferation and invasion were suppressed in the Sh-HULC group, while miR-377-5p showed the opposite. Further experiments exhibited that miR-377-5p was targeted by HULC, and an negative correlation between HULC and miR-377-5p was observed. Importantly, the in vivo experiments indicated that down-regulation of HULC could inhibit tumor growth. Taken together, our research demonstrated that down-regulation of HULC plays an anti-cancer role through restrainingHepG2 cell proliferation and invasion.Conclusions: In summary, our in vitro and in vivo findings confirmed HULC to play a role in the progression of HCC, with the underlying mechanism possibly involving the miR-377-5p/HIF-1α pathway.

The long and short non-coding RNAs modulating EZH2 signaling in cancer

“…It is noteworthy that most experiments have focused on revealing the role of tumor-promoting factors. It has been shown that siRNA and shRNA can be applied to down-regulate lncRNAs [ 410 , 411 ]. Thus, next experiments can focus on the targeting of ncRNAs and translating these findings in clinical settings.…”

Epigenetic Changes Affecting the Development of Hepatocellular Carcinoma

“…HULC knockdown increased the chemosensitivity to oxaliplatin through the repression of cell growth and the acceleration apoptosis in HCC cells. Further analysis confirmed its mode of action through the regulation of the miR-383-5p/vesicle-associated membrane protein-2 axis, the miR-377-5p/HIF-1α pathway, and the miR-134-5p/FOXM1 axis [41,42]. Emerging evidence provides new insights on the role of other lncRNAs in HCC development, such as NEAT1, ANRIL, SNHG1, or H19 [36,69].…”

“…Regulation of mitochondrial metabolism [38] HULC Growth of liver cancer stem cells [39] Chemosensitivity of anti-cancer drug oxaliplatin inhibition [40] Regulation of miR-383-5p/vesicle-associated membrane protein-2 pathway; miR-377-5p/HIF-1α pathway and miR-134-5p/FOXM1 pathway [41,42] On the contrary, hypermethylation may disturb the function of another group of genes. One of the signaling pathways affected by hypermethylation is the WNT/β-catenin signaling.…”

Dexmedetomidine reduces propofol-induced hippocampal neuron injury by modulating the miR-377-5p/Arc pathway

“…Moreover, miR-377-5p overexpression aggrandized myocardial dysfunction as well as apoptosis, and promoted the release of inflammatory factors [ 81 ]. However, miR-377-5p downregulation suppressed the proliferation and invasion of HepG2 cells belonging to hepatocellular carcinoma cell lines [ 82 ]. Interestingly, Li Y, et al [ 83 ] has demonstrated that miR-377-5p was up-regulated after propofol treatment and contributed to induce neurotoxicity, the result was consistent with our finding.…”