Effect of TRIM8 gene on proliferation and invasion of hepatocellular carcinoma cells and its mechanism

doi:10.3877/cma.j.issn.2095-3232.2019.05.018

Abstract

Abstract:

Objective

To evaluate the effect of tripartite motif 8 (TRIM8) gene on the proliferation and invasion of hepatocellular carcinoma cells and the underlying mechanism.

Methods

Lentiviral vectors expressing TRIM8 were synthesized and hepatocellular carcinoma cell lines Hep3B-TRIM8 group which stably expressed TRIM8, Huh-7-TRIM8 over-expression groups and control cell lines of Hep3B-Vector, Huh-7-Vector (control groups) were constructed. The expression level of TRIM8 in hepatocellular carcinoma cells was determined by RT-PCR. The proliferation ability of hepatocellular carcinoma cells in two groups was evaluated by CCK-8 assay. The clone formation ability was assessed by plate colony formation assay. The invasion ability was determined by Transwell assay. The expression levels of TRIM8 and P53 proteins were quantitatively measured by Western blot and immunohistochemistry. The experimental data were statistically compared by t test.

Results

The expression levels of TRIM8 mRNA in the Hep3B-TRIM8 and Huh-7-TRIM8 over-expression groups were (4.47±0.29)×10^-3 and (2.43±0.41)×10^-3 respectively, significantly higher than (0.60±0.10)×10^-3 and (0.19±0.01)×10^-3 in the control groups (t=12.570, 5.458; P<0.05). The cell growth curve demonstrated that the proliferation ability of hepatocellular carcinoma cells in the over-expression groups were significantly weaker compared with those in the control groups. The number of cell colony in the Hep3B-TRIM8 and Huh-7-TRIM8 over-expression groups was 69±9 and 52±8 respectively, significantly less than 132±11 and 103±5 in the control groups (t=-4.503, -5.643; P<0.05). In the Hep3B-TRIM8 and Huh-7-TRIM8 over-expression groups, the number of trans-membrane cells was 15.7±2.9 and 11.3±1.5 respectively, remarkably lower than 42.3±3.2 in the control group (t=-6.191, -11.360; P<0.05). The expression levels of TRIM8 and P53 proteins in the over-expression groups were significantly up-regulated compared with those in the control groups.

Conclusions

TRIM8 gene can inhibit the proliferation and invasion of hepatocellular carcinoma cells, probably by restoring the anti-cancer activity of P53 and synergistically exerting anti-cancer effect.

Key words: Carcinoma, hepatocellular, Cell proliferation, Neoplasm invasiveness, Tumor suppressor protein p53

Huiling Zhang, Jiaojiao Gu, Siyuan Hong, Xiaolan Lin. Effect of TRIM8 gene on proliferation and invasion of hepatocellular carcinoma cells and its mechanism[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2019, 08(05): 458-462.

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References 18

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