MiR‐490‐5p inhibits the stemness of hepatocellular carcinoma cells by targeting ECT2

Male 0303 health sciences Binding Sites Carcinoma, Hepatocellular Cell Survival Liver Neoplasms Hep G2 Cells Middle Aged Prognosis Disease-Free Survival 3. Good health Gene Expression Regulation, Neoplastic MicroRNAs 03 medical and health sciences Hyaluronan Receptors Ki-67 Antigen Proto-Oncogene Proteins Humans Female AC133 Antigen 3' Untranslated Regions Aged Cell Proliferation
DOI: 10.1002/jcb.27459 Publication Date: 2018-09-12T06:53:18Z
ABSTRACT
AbstractTo explore the targeting relationship between miR‐490‐5p and ECT2 in hepatocellular carcinoma (HCC) and the influences of miR‐490‐5p and ECT2 on the stemness of HCC cells. The expressions of miR‐490‐5p and ECT2 in HCC tissues and adjacent tissues were identified by quantitative real‐time polymerase chain reaction (qRT‐PCR). The relationships between the expression levels of miR‐490‐5p/ ECT2 and the overall/disease‐free survival (OS/DFS) of patients with HCC were evaluated using correlative curves. In addition, the targeting relationship between miR‐490‐5p and ECT2 was predicted by TargetScan and verified by dual‐luciferase reporter assay. Plasmid transfection was used for overexpression of ECT2 in HepG2 cells, and transfection efficiency was verified by qRT‐PCR. Cell Counting Kit‐8 assay and cell sphere‐formation assay were conducted to detect the proliferation and sphere‐formation ability of HCC cells, respectively. Cell populations with different cell transfections were sorted using flow cytometry. The expression levels of proteins in the stem cell signaling pathway were determined using Western blot analysis. MiR‐490‐5p was remarkably downregulated, yet ECT2 was upregulated in HCC tissues compared with adjacent tissues. MiR‐490‐5p expression was positively correlated with OS and DFS of patients with HCC, which were otherwise negatively correlated with ECT2 expression. ECT2 was validated to be the downstream target of miR‐490‐5p. Overexpression of miR‐490‐5p restrained the sphere formation ability, stemness, and proliferation of HCC cells. MiR‐490‐5p repressed the stemness of HCC cells through inhibiting the expression of ECT2. MiR‐490‐5p may be an underlying therapeutic target in HCC treatment.
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