激活FXR抑制结肠癌细胞浸润转移及MMP-7的表达

目的:探讨法尼酯X受体(FXR)特异性激动剂GW4064抑制结肠癌细胞浸润转移的机制。方法:在体外培养人结肠癌细胞HT-29,应用GW4064作用于结肠癌细胞,以四唑氮蓝还原法(MTT)检测细胞活性的变化。用transwell小室研究结肠癌细胞的迁移及浸润。用RT-PCR检测FXRm RNA及MMP-7mRNA表达的变化,用western blot检测FXR及MMP-7蛋白表达的变化。结果:MTT结果显示GW4064作用于人结直肠HT-29细胞的生长抑制率呈浓度依赖性;transwell小室结果显示GW4064抑制结肠癌细胞的浸润转移,与对照组相比,差异具有统计学意义(P0.05),RT-PCR及Western blot显示GW4064促进FXR m RNA及蛋白表达,抑制MMP-7mRNA及蛋白的表达,与对照组相比差异有统计学意义(P0.05)。结论:GW4064抑制结肠癌细胞的生长及转移,上调HT-29细胞FXR m RNA及蛋白的表达,下调HT-29细胞MMP-7 m RNA及蛋白的表达。FXR被激活后抑制结肠癌细胞转移,MMP-7可能是其作用通路之一。

Activation of Farnesoid X Receptor Inhibited the Infiltration and Metastasis and MMP-7's Expression of Colon Cancer Cells

ABSTRACT Objective: To investigate the mechanism of the farnesoid X receptor (FXR) activation by GW4064 to inhibit the growth of colon cancer cells. Methods: The effects of specific FXR agonist (GW4064) on the growth of HT-29 cells were studied in vitro by MTT. The infiltration and metastasis of HT-29 cells were detected by the transwell inserts. The mRNA expression of FXR and MMP-7 were determined by RT-PCR, The protein expression of FXR and MMP-7 were measured by Western blot. Results: MTT results showed that the growth inhibition rate of GW4064 in human colorectal HT-29 cells was concentration dependent; Transwell inserts re- sults showed that GW4064 inhibited the invasion and metastasis of colon cancer cells. Compared with the control group, the difference was statistically significant (P<0.05). RT-PCR and Western blot display GW4064 promoted FXR mRNA and protein expression and inhi- bited the expression ofMMP-7. The expression of mRNA and protein was significantly different from that of the control group (P<0.05). Conclusion: GW4064 inhibited metastasis of colon cancer cells. The expression of mRNA and protein of FXR was up-regulated by FXR specific agonist in cell line HT-29, and MMP-7 is just opposite. The activation of FXR may inhibit the metastasis of colon cancer cells by inhibiting MMP-7.