The Effect Of Janus Kinase Inhibitor AG490on Invasion And Metastasis Potential Of Human Hepatoma Cell SMMC-7721

Objective To investigate the effect of Janus kinase (JAK) inhibitorAG490on invasion and metastasis potential of human hepatoma cell lineSMMC-7721, and to explore the role of JAK-STAT3signaling pathway inregulating the invasion of liver cancer.Methods1. JAK inhibitor AG490was added into the culture mediafor SMMC-7721cells, the inhibiting effect of AG490on hepatocarcinomawas assayed by MTT.2. SMMC-7721cells were treated with AG490at theconcentration of5μmol/L and10μmol/L, at which concentration theinhibiting effect of AG490on proliferation of SMMC-7721cells were notsignificant. The effect of AG490on invasion and metastatic ability ofhepatocarcinoma was evaluated by Transwell chamber.3.48h after theadministration of AG490at the final concentration5μmol/L and10μmol/L,RT-PCR was performed to detect mRNA expression of the MMP-2andVEGF. Western-blot was conducted to detect protein expression andactivation of STAT3, MMP-2and VEGF.Results48hours after the administration of Janus kinase inhibitor AG490, migrant assay demonstrated that the number of LoVo cells incontrol group is larger than those in5μmol/L and10μmol/L AG490group (120.2±13.9vs81.4±8.6vs54.4±6.7, P<0.05), showing that themigration potential of SMMC-7721cells decreased significantly.Transwell invasion assay suggested that the LoVo cells in control groupwas the largest, followed by those in5μmol/L and10μmol/L AG490group (74.6±6.3vs58.2±7.2vs38.2±5.7, P<0.05), thus the invasioncapability of SMMC-7721cells decreased significantly. RT-PCR analysissuggested that the expression of MMP-2mRNA and VEGF mRNAdropped down. The results of western-blot showed that the activation ofSTAT3was suppressed, the expression of MMP-2and VEGF was reduced(P<0.05).Conclusion1.AG490can inhibit the growth and decrease theproliferation of hepatocarcinoma SMMC-7721cells, and the inhibitioneffect increase along with concentration growing and time progressing.2.Metastasis and invasion potential of hepatocarcinoma SMMC-7721cellswas reduced after being treated with AG490.3. Down-regulation of thelevel of MMP-2and VEGF expression is one of the possible mechanismthat mediated the decreasing of the invasion ability of SMMC-7721.4.Blocking the activation of STAT3can reduce the expression level ofMMP-2and VEGF and suppress the invasion ability of SMMC-7721cellsin vitro.