Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells
Authors Chen GS, Zhou N, Li J, Li T, Zhang Z, Si Z
Received 22 September 2015
Accepted for publication 24 December 2015
Published 27 May 2016 Volume 2016:9 Pages 3067—3076
Checked for plagiarism Yes
Review by Single-blind
Peer reviewers approved by Dr Federico Perche
Peer reviewer comments 2
Editor who approved publication: Professor Daniele Santini
Guang Shun Chen, Ning Zhou, Jie-Qun Li, Ting Li, Zhong-Qiang Zhang, Zhong-Zhou Si
Department of Organ Transplantation and General Surgery, The Second Xiangya Hospital, Central South University, Changsha, People’s Republic of China
Objective: To study microRNA (miR)-20a expression in hepatocellular carcinoma (HCC) and its effects on the proliferation, migration, and invasion of HepG2.
Methods: The real-time polymerase chain reaction was used to detect the expression of miR-20a in HCC tissue and normal tissue, as well as in HCC cell lines and normal liver cells. miR-20a mimic and miR negative control (NC) were transfected into HepG2 cells. MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay was used to detect cell proliferation. Annexin fluorescein isothiocyanate/propidium iodide assay was run to examine the early apoptosis of cells. Transwell chamber assay was carried out to investigate the cell invasion and migration abilities.
Results: miR-20a was lowly expressed both in HCC tissues and HCC cell lines. After transfection of exogenous miR-20 mimics, miR-20a expression in HepG2 cells was significantly increased by 61.29% compared to the blank group (P<0.01). MTT assay showed that the growth of HepG2 cells in the miR-20a mimics group was significantly inhibited, and optical density values during the 36–96 hour time period were dramatically decreased compared to the blank group (P<0.01). Apoptosis rates of the miR-20a mimics group were higher than those of the blank and NC groups (both P<0.01). The number of HCC cells after transfection by miR-20a mimics in the G1 and S phases were 15.88% and 7.89%, respectively, which were lower than in the blank and NC groups (both P<0.05). Transwell assay showed that in the miR-20a mimics group the number of cell migration and invasion were 0.459 and 0.501 times that of the blank group (both P<0.01), and the migration and inhibition rates were 54.1% and 51.4%, respectively. After closing target gene CCND1 in HepG2 cells, the number of cell migration and invasion in the small interfering (si)-CCND1 group were 0.444 and 0.435 times that of the si-NC group (P<0.05); and compared to the si-NC group, the migration and inhibition rates were 55.6% and 56.5%, respectively.
Conclusion: miR-20a can inhibit the growth, invasion, and migration of HepG2 cells, and is therefore promising as a new molecular target for diagnosis and therapy of HCC.
Keywords: miR-20a, HepG2 cells, proliferation, migration, invasion
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