2019OncoTargets and TherapyOpen access

Overexpression of miR-450 affects the biological behavior of HepG2 cells by targeting DNMT3a

Yan Wang, Lina Wang, Xue Yu, Jianping Duan

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Abstract

PURPOSE: This study was designed to explore the regulation mechanism of miR-450 in the development of hepatocarcinoma, and the effects of overexpression of miR-450 on biological behaviors such as proliferation, migration, and invasion of hepatoma cells. METHODS: HepG2 cells were divided into miR-450 mimics group, miR-450 inhibitor group, miR-450 mimics NC group, miR-450 inhibitor NC group, and blank group. MTT assay was served to measure cell proliferation, and Transwell assay was used to test cell migration and invasion. Additionally, cell cycle was detected by flow cytometry and apoptosis was examined with AnnexinV-PI double staining. After the target gene of miR-450 was predicted by bioinformatics software, Western blot and dual luciferase reporter gene experiment were applied to verify the relationship between miR-450 and target gene. RESULTS: The MTT and Transwell assay indicated that overexpression of miR-450 inhibited the proliferation, invasion, and migration of HepG2 cells. The flow cytometry analysis showed that overexpression of miR-450 arrested the cell cycle in the G1 phase. Meanwhile, Annexin V-PI double staining assay revealed that overexpression of miR-450 promoted apoptosis of HepG2 cells. However, silencing miR-450 in HepG2 cells promoted proliferation and invasion, and reduced apoptosis. Moreover, we found that DNMT3a was the target gene of miR-450. CONCLUSIONS: miR-450 could inhibit proliferation, invasion, and migration via regulating DNMT3a in hepatocarcinoma cells, which provided a theoretical basis for the treatment of liver cancer.

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PURPOSE: This study was designed to explore the regulation mechanism of miR-450 in the development of hepatocarcinoma, and the effects of overexpression of miR-450 on biological behaviors such as proliferation, migration, and invasion of hepatoma cells. METHODS: HepG2 cells were divided into miR-450 mimics group, miR-450 inhibitor group, miR-450 mimics NC group, miR-450 inhibitor NC group, and blank group. MTT assay was served to measure cell proliferation, and Transwell assay was used to test cell migration and invasion. Additionally, cell cycle was detected by flow cytometry and apoptosis was examined with AnnexinV-PI double staining. After the target gene of miR-450 was predicted by bioinformatics software, Western blot and dual luciferase reporter gene experiment were applied to verify the relationship between miR-450 and target gene. RESULTS: The MTT and Transwell assay indicated that overexpression of miR-450 inhibited the proliferation, invasion, and migration of HepG2 cells. The flow cytometry analysis showed that overexpression of miR-450 arrested the cell cycle in the G1 phase. Meanwhile, Annexin V-PI double staining assay revealed that overexpression of miR-450 promoted apoptosis of HepG2 cells. However, silencing miR-450 in HepG2 cells promoted proliferation and invasion, and reduced apoptosis. Moreover, we found that DNMT3a was the target gene of miR-450. CONCLUSIONS: miR-450 could inhibit proliferation, invasion, and migration via regulating DNMT3a in hepatocarcinoma cells, which provided a theoretical basis for the treatment of liver cancer.

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Available abstract

PURPOSE: This study was designed to explore the regulation mechanism of miR-450 in the development of hepatocarcinoma, and the effects of overexpression of miR-450 on biological behaviors such as proliferation, migration, and invasion of hepatoma cells. METHODS: HepG2 cells were divided into miR-450 mimics group, miR-450 inhibitor group, miR-450 mimics NC group, miR-450 inhibitor NC group, and blank group. MTT assay was served to measure cell proliferation, and Transwell assay was used to test cell migration and invasion. Additionally, cell cycle was detected by flow cytometry and apoptosis was examined with AnnexinV-PI double staining. After the target gene of miR-450 was predicted by bioinformatics software, Western blot and dual luciferase reporter gene experiment were applied to verify the relationship between miR-450 and target gene. RESULTS: The MTT and Transwell assay indicated that overexpression of miR-450 inhibited the proliferation, invasion, and migration of HepG2 cells. The flow cytometry analysis showed that overexpression of miR-450 arrested the cell cycle in the G1 phase. Meanwhile, Annexin V-PI double staining assay revealed that overexpression of miR-450 promoted apoptosis of HepG2 cells. However, silencing miR-450 in HepG2 cells promoted proliferation and invasion, and reduced apoptosis. Moreover, we found that DNMT3a was the target gene of miR-450. CONCLUSIONS: miR-450 could inhibit proliferation, invasion, and migration via regulating DNMT3a in hepatocarcinoma cells, which provided a theoretical basis for the treatment of liver cancer.

Key concepts: Cancer research, Chemistry, Cell biology, Pharmacology, Molecular biology, Computational biology, Biology

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