IL-32γ induces the expression of MMP2 in hepatic stellate cells LX-2 via activating Akt signaling pathway

doi:10.3877/cma.j.issn.2095-3232.2017.04.021

Abstract

Abstract:

Objective

To investigate the expression of matrix metalloproteinase 2 (MMP2) in hepatic stellate cells LX-2 induced by IL-32γ and its mechanism.

Methods

Different concentrations (0, 10, 20 μg/L) of IL-32γ protein were co-cultured with LX-2 cells, and the expression level of MMP2 mRNA, MMP2 protein and activated Akt was respectively detected by RT-PCR, ELISA and Human Phospho-Akt Immunoassay kit. Different concentrations (0, 25, 50 μg/L) of recombinant human activated Akt protein were respectively co-cultured with LX-2 cells, and different concentrations (0, 10, 20 μmol/L) of Akt inhibitor Perifosine were respectively co-cultured with IL-32γ and LX-2 cells, and the expression of MMP2 protein was detected by ELISA. The expression levels of Akt, MMP2 mRNA and MMP2 protein among three groups were compared using one-way analysis of variance and LSD-t test.

Results

After the co-culture of different concentrations of IL-32γ protein and LX-2 cells, the expression levels of MMP2 mRNA and MMP2 protein were respectively (0.9±0.4), (5.5±1.4), (12.2±2.6) and (53±13), (234±38), (523±101) μg/L, and appeared as dose-dependent elevation (LSD-t=5.517, 3.952 and 7.855, 4.634; P<0.05). The expression of Akt in LX-2 cells could be induced by IL-32γ, the levels was repectively (86±9), (287±60) and (650±170) RFUs in accordance with the concetrations, which appeared as dose-dependent elevation (LSD-t=5.700, 3.489; P<0.05). The expression of MMP2 in LX-2 cells could be induced by recombinant human activated Akt protein, the levels was repectively (54±14), (277±45) and (550±132) μg/L in accordance with the concetrations, which appeared as dose-dependent elevation (LSD-t=8.155, 3.387; P<0.05). Different concentrations of Perifosine could inhibit the inducing effect of IL-32γ upon the expression of MMP2 in LX-2 cells, the expression level of MMP2 protein was respectively (553±86), (233±28) and (89±12) μg/L in accordance with the concetrations, which appeared as dose-dependent reduction (LSD-t=6.130, 8.083; P<0.05).

Conclusions

IL-32γ may induce the expression of MMP2 in hepatic stellate cells LX-2 via activating the Akt signaling pathway, thereby participating in the incidence and metastasis of hepatic fibrosis and liver cancer.

Key words: Interleukins, Matrix metalloproteinase 2, LX-2 cells

Meilian Liang, Xiaoyan Chen, Fan Zhang, Xingfei Pan. IL-32γ induces the expression of MMP2 in hepatic stellate cells LX-2 via activating Akt signaling pathway[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2017, 06(04): 328-331.

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References 20

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IL-32γ蛋白浓度（μg/L）	MMP2 mRNA	MMP2蛋白（μg/L）
20	12.2±2.6	523±101
10	05.5±1.4	234±380
0	00.9±0.4	053±130
F	33.179	42.729
P	<0.05	0<0.05

IL-32γ蛋白浓度（μg/L）	MMP2 mRNA	MMP2蛋白（μg/L）
20	12.2±2.6	523±101
10	05.5±1.4	234±380
0	00.9±0.4	053±130
F	33.179	42.729
P	<0.05	0<0.05

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