2018IUBMB LifeOpen access

MiR‐490‐3p inhibits autophagy via targeting ATG7 in hepatocellular carcinoma

Yingliang Ou, Jiafa He, Yubin Liu

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Abstract

The miR-490-3p was transfected into HepG2 cells to explore the correlation between miR-490-3p and hepatocellular carcinoma cell proliferation, apoptosis, and autophagy and its downstream target gene ATG7. Then we could possibly provide a mechanism for the treatment of hepatocellular carcinoma. MiR-490-3p was screened out by fold change > 4 and P < 0.01 using gene microarray data. The expression level of miR-490-3p was tested by qRT-PCR and the prognosis analysis was achieved by using TCGA data. The cell proliferation was tested via colony formation assay and CCK-8 after the miR-490-3p mimics were transfected into HepG2 cells; the variations of cell cycle and apoptosis was examined by flow cytometry assay; the number of autophagosome was observed by electron microscopy and the changes of autophagy-relative protein LC-II and LC-I as well as their ratio was tested by western blot. MiR-490-3p is low expressed in hepatocellular carcinoma cell lines and tissues. The results of TCGA showed that miR-490-3p high expression indicated better prognosis. After HepG2 cells were transfected with miR-490-3p mimics, cell viability was increased, cell proliferation was enhanced, cell cycle was blocked in G0/G1 phase, cell apoptosis rate was increased, the number of autophagosomes was reduced, autophagy-associated protein LC-II was decreased, and LC-I was increased and their ratio was decreased. After 3-MA was added, cell proliferation was declined, cell apoptosis rate was increased. Besides, the autophagy was inhibited by knocking down the ATG7, which promoted the cell apoptosis. MiR-490-3p could suppress cell proliferation, retard cell cycle and upgrade cell apoptosis by inhibiting autophagy in HCC cells via targeting ATG7. © 2018 IUBMB Life, 70(6):468-478, 2018.

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The miR-490-3p was transfected into HepG2 cells to explore the correlation between miR-490-3p and hepatocellular carcinoma cell proliferation, apoptosis, and autophagy and its downstream target gene ATG7. Then we could possibly provide a mechanism for the treatment of hepatocellular carcinoma. MiR-490-3p was screened out by fold change > 4 and P < 0.01 using gene microarray data. The expression level of miR-490-3p was tested by qRT-PCR and the prognosis analysis was achieved by using TCGA data. The cell proliferation was tested via colony formation assay and CCK-8 after the miR-490-3p mimics were transfected into HepG2 cells; the variations of cell cycle and apoptosis was examined by flow cytometry assay; the number of autophagosome was observed by electron microscopy and the changes of autophagy-relative protein LC-II and LC-I as well as their ratio was tested by western blot. MiR-490-3p is low expressed in hepatocellular carcinoma cell lines and tissues. The results of TCGA showed that miR-490-3p high expression indicated better prognosis. After HepG2 cells were transfected with miR-490-3p mimics, cell viability was increased, cell proliferation was enhanced, cell cycle was blocked in G0/G1 phase, cell apoptosis rate was increased, the number of autophagosomes was reduced, autophagy-associated protein LC-II was decreased, and LC-I was increased and their ratio was decreased. After 3-MA was added, cell proliferation was declined, cell apoptosis rate was increased. Besides, the autophagy was inhibited by knocking down the ATG7, which promoted the cell apoptosis. MiR-490-3p could suppress cell proliferation, retard cell cycle and upgrade cell apoptosis by inhibiting autophagy in HCC cells via targeting ATG7. © 2018 IUBMB Life, 70(6):468-478, 2018.

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Available abstract

The miR-490-3p was transfected into HepG2 cells to explore the correlation between miR-490-3p and hepatocellular carcinoma cell proliferation, apoptosis, and autophagy and its downstream target gene ATG7. Then we could possibly provide a mechanism for the treatment of hepatocellular carcinoma. MiR-490-3p was screened out by fold change > 4 and P < 0.01 using gene microarray data. The expression level of miR-490-3p was tested by qRT-PCR and the prognosis analysis was achieved by using TCGA data. The cell proliferation was tested via colony formation assay and CCK-8 after the miR-490-3p mimics were transfected into HepG2 cells; the variations of cell cycle and apoptosis was examined by flow cytometry assay; the number of autophagosome was observed by electron microscopy and the changes of autophagy-relative protein LC-II and LC-I as well as their ratio was tested by western blot. MiR-490-3p is low expressed in hepatocellular carcinoma cell lines and tissues. The results of TCGA showed that miR-490-3p high expression indicated better prognosis. After HepG2 cells were transfected with miR-490-3p mimics, cell viability was increased, cell proliferation was enhanced, cell cycle was blocked in G0/G1 phase, cell apoptosis rate was increased, the number of autophagosomes was reduced, autophagy-associated protein LC-II was decreased, and LC-I was increased and their ratio was decreased. After 3-MA was added, cell proliferation was declined, cell apoptosis rate was increased. Besides, the autophagy was inhibited by knocking down the ATG7, which promoted the cell apoptosis. MiR-490-3p could suppress cell proliferation, retard cell cycle and upgrade cell apoptosis by inhibiting autophagy in HCC cells via targeting ATG7. © 2018 IUBMB Life, 70(6):468-478, 2018.

Key concepts: Autophagy, Apoptosis, Cell growth, Cell cycle, Flow cytometry, Transfection, Hepatocellular carcinoma, Cell

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