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Objective To investigate the effects of inhibiting KDM2A gene on the proliferation, invasion and migration of liver cancer cells and its possible regulatory mechanism. Methods Forty pairs of HCC tissues and their adjacent normal counterparts were collected from 2014 to 2017 in Tongji Hospital, Tongji Medical College Affiliated to Huazhong University of Science and Technology. Human liver cancer cell lines HepG2, Huh7, HCCLM3, MHCC-97H and normal liver cells LO2 were cultured in vitro. The mRNA and protein expression levels of KDM2A in HCC tissues and cells were detected by qRT-PCR and Western blotting. Huh7 cells were taken and set up as follows: (1) si-NC group (transfected with si-NC) and si-KDM2A group (transfected with si-KDM2A); (2) mimic-NC group (transfected with mimic-NC), miRNA-29a-3p mimic group (transfected with miRNA-29a-3p mimic), inhibitor-NC group (transfected with inhibitor-NC) and miRNA-29a-3p inhibitor group (transfected with miRNA-29a-3p inhibitor). The mRNA and protein expression levels of KDM2A were detected by qRT-PCR and Western blotting. The invasion and migration of cells were detected by Transwell, the proliferation of cells was detected by CCK-8 methods. The online databases TargetScan, miRDIP, miRWalk, Starbase and miRDB were used to predict the binding sites of KDM2A and miR-29a-3p. The KDM2A 3'-UTR (WT) or KDM2A 3'-UTR (MUT) report plasmid was co-transfected with NC-miRNA or miR-29a-3p mimics respectively for 48 h in 293T cells, and the luciferase activity was detected by the luciferase reporter gene detection system. Results Compared with adjacent normal counterparts, the relative mRNA and protein expression levels of KDM2A in HCC tissues increased significantly (P<0.05). Compared with LO2, the relative mRNA and protein expression levels of KDM2A in HepG2, Huh7, HCCLM3 and MHCC-97H increased significantly (P<0.05). Compared with si-NC group, the proliferation, invasion and migration of Huh7 cells in si-KDM2A group decreased significantly (P<0.05 or P<0.01). The analysis results of TargetScan, miRDIP, miRWalk, Starbase and miRDB showed that there were binding sites between KDM2A and miR-29a-3p. The results of the dual luciferase reporter assay showed that miR-29a-3p mimic significantly reduced KDM2A-MUT luciferase activity (P<0.01). After overexpression of miRNA-29a-3p, the relative mRNA and protein expression levels of KDM2A were decreased (P<0.01), the proliferation, invasion and migration abilities were decreased (P<0.05) in Huh7 cells. After inhibiting the expression of miRNA-29a-3p, the relative mRNA and protein expression levels of KDM2A were increased (P<0.05), the proliferation, invasion and migration abilities were enhanced (P<0.05) in Huh7 cells. Conclusion Inhibiting the expression of KDM2A can reduce the proliferation and migration ability of Huh7 cells. miR-29a-3p may be the upstream regulator of KDM2A and participate in the occurrence and development of hepatocellular carcinoma.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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