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| The Effects of miR-490-3p on Proliferation and Migration of HeLa Cells |
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Abstract microRNA-490-3p (miR-490-3p) is down-regulated and may act as a tumor inhibitor in many tumors, such as gastric cancer, ovarian carcinoma, endometrial carcinoma and so on. However, specific roles of miR-490-3p are not well understood in cancers. This study aims to investigate the effects of miR-490-3p on the proliferation and metastasis of human (Homo sapiens) cervical tumor cells HeLa. In order to investigate the effects of miR-490-3p on proliferation, migration, cell cycle and EMT (epithelial to mesenchymal transition) of HeLa cells, miR-NC mimic or miR-490-3p mimic was transfected into HeLa cells. The expression level of miR-490-3p was detected using qRT-PCR, then, cell counting was used to measure the cell proliferation, and wound healing assay was applied to detect the ability of migration. Moreover, cell cycle was assayed by flow cytometry, and EMT proteins were detected by Western blot, including E-cadherin, β-catenin, vimentin and glycogen synthase kinase 3β (GSK-3β). The results showed that, in comparison with negative controls, the expression level of miR-490-3p was significantly improved by miR-490-3p mimic transfection (P＜0.01), which showed that miR-490-3p mimic was effctive to express miR-490-3p. Then, the HeLa cells transfected with miR-490-3p mimic exhibited signicantly slower growth than negative controls (P<0.05), which indicated that the overexpression of miR-490-3p reduced cell proliferation. Wound healing assay showed that, after transfected with miR-490-3p mimic, HeLa cells migration was reduced (P＜0.01), which suggested miR-490-3p transfection inhibited migration ability of HeLa cells. Flow cytometry analysis confirmed that, comparing with control group, the proportion of HeLa cells transfected with miR-490-3p in G2/M phase displayed a significant increase (P<0.01). This induced cell cycle arresting at G2/M phase. miR-490-3p probably inhibited cells growth through arresting cell cycle of HeLa cells. Western blot showed that the EMT proteins were also regulated through transfection with miR-490-3p into HeLa cells, the introduction of exogenous miR-490-3p was followed by an increase in expression of E-cadherin (P＜0.05) which was a biomarker of epithelial cells and decreases in β-catenin (P＜0.01), vimentin (P＜0.05) and GSK-3β (P＜0.01) expression. The result showed that the EMT of HeLa cells was inhibited and indicated that miR-490-3p may inhibit cell invasiveness by repressing EMT, which was a key mechanism in cancer cell invasiveness and metastasis. Collectively, these results displayed the overexpression of miR-490-3p reduced cell proliferation and migration and promoted G2/M phase arrest, which suggested that miR-490-3p may act as a tumor suppressor gene in HeLa cells. These data indicates that miR-490-3p can be used as a potential therapeutic target and a molecular marker for cancer prediction in human cervical cancer.
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Received: 12 April 2017
Published: 30 October 2017
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