Leptin promotes the proliferation and invasion of human hepatocellular carcinoma cell line Huh-7 in vitro / 肿瘤

ABSTRACT

Objective: To observe the effect of leptin on proliferation and invasion of human hepatocellular carcinoma cell line Huh-7 and explore its possible molecular mechanism. Methods: Huh-7 cells were cultured and treated with leptin at different concentrations of 5, 10, 50, 100 and 200 ng/mL for 12, 24 and 48 h. Huh-7 cells without leptin treatment were used as control group. The proliferation rate of the Huh-7 cells was measured by MTT assay. The change of cell cycle was detected by flow cytometry analysis. Transwell chamber assay was performed to determine the effect of leptin 100 ng/mL on the invasion capability of Huh-7 cells 24 h later. The effect of leptin on mRNA expression of matrix metalloproteinase (MMP) of Huh-7 cells was detected by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). Results: Leptin significantly promoted the proliferation of Huh-7 cells. The effect was dose-dependent in certain degree. Flow cytometry analysis found that leptin significantly induced decrease in proportion of cells in G0/1 phase and significantly increased cell proportion in S phase. The invasion capability of Huh-7 cells was significantly increased after leptin 10 ng/mL treatment for 24 h (P <0.01). The number of cells penetrated polycarbonate membrane were (41.80 ± 3.19) and (18.20 ± 2.86), respectively. Real time fluorogentic quantitative PCR analysis revealed that the expression level of MMP-2 and MMP-9 mRNA expression were 4.25-fold and 2.73-fold as the control group (P <0.01), respectively. Conclusion: Leptin significantly increases the proliferation and invasion capabilities of Huh-7 cells in vitro. The possible mechanism may be related with up-regulation of MMPs level.