Authors

Dianyan Chen*, Sandu Liu*, #, Huaping Huang**

Departments

*Department of Hepatology, The People's Hospital of Qiannan, Duyun, PR China - **Department of Respiratory Medicine, First Affiliated Hospital of Hainan Medical College, Haikou, PR China

Abstract

Objective: To investigate the inhibitory effect of LAP1 overexpression on proliferation, migration and invasion of hepatocellular carcinoma cells.

Methods: The study included 3 × 106 293T cells which were cultured in a DMEM medium for 24 hours and transfected when the cell density reached about 75%. In the sterile centrifugal tube, 1 ml of the DMEM medium was added, and the skeleton plasmids mCherry-IRES-GFP, packaging plasmids psPAX 2 and pMD2G were fully mixed at a ratio of 4:2:1. Polyethyleneimine (PEI) was added with 21 μL, then blended and placed stilly at room temperature for 15 minutes. We collected supernatant containing lentivirus, and HuH-7 human hepatocellular carcinoma cell lines were inoculated into six well plates with a fusion degree of about 35%. Each hole was shaken well by adding 150 mu L virus suspension. The culture medium was replaced about ten hours after incubation. Non-infected cells were set as a control group. The expression of LAP1 in hepatocellular carcinoma cells and normal adjacent tissues was observed by immunohistochemistry. The effects of LAP1 overexpression on proliferation, cell cycle, cell migration and invasion of hepatocellular carcinoma cells were observed.

Results: The expression of LAP1 in hepatocellular carcinoma was significantly lower than that in adjacent normal tissues. With the increase of time, the cell proliferation ability of both groups showed an increasing trend, but from the third day, the cell proliferation ability of the LAP1 overexpression group was significantly lower than that of the control group (P<0.05). Compared with the control group, the cloning ability of Hun7 cells in the LAP1 overexpression group was significantly lower (P<0.05). The blockade of the Hun7G1/G0 phase in the LAP1 overexpression group was significantly higher than that in the control group (P<0.05). Compared with the control group, the migration ability of Hun7 cells in the LAP1 overexpression group was significantly decreased (P<0.05). The invasive ability of the Hun7 cells in the LAP1 overexpression group was significantly lower than that in the control group (P<0.05).

Conclusion: The overexpression of LAP1 in hepatocellular carcinoma cells can significantly reduce the proliferation, migration, and invasion of hepatocellular carcinoma cells, induce the senescence of hepatocellular carcinoma cells, and then inhibit the development of tumours.

Keywords

LAP1, overexpression, hepatocellular carcinoma cells, proliferation, migration, invasion, inhibition.

DOI:

10.19193/0393-6384_2020_3_328