PPAR-γ经其配体激活后对胆管癌细胞侵袭力的影响及相关基因表达的调控

Effect of Ligand Pioglitazone Activating PPAR-γ on the Invasiveness of Cholangiocarcinoma Cell in vitro and the Expression Regulation of Related Genes

目的探讨过氧化物酶体增殖物激活受体-γ(PPAR-γ)经其配体吡咯列酮(pioglitazone,PGZ)激活后,对胆管癌细胞体外侵袭力的影响及机制。方法体外培养人肝门胆管癌细胞系QBC939细胞。实验分为实验组(又分为不同PGZ浓度组)和对照组(无PGZ干预);MTT法检测PGZ对QBC939细胞的增殖抑制率;荧光定量PCR(FQ-PCR)检测PGZ对QBC939细胞基质金属蛋白酶-7(MMP-7)、金属蛋白酶组织抑制剂-1(TIMP-1)表达的影响;Matrigel侵袭实验和过河实验检测PGZ对QBC939细胞体外侵袭力和运动能力的影响。结果MTT结果显示干预12h、其PGZ5～40μmol/L时,细胞毒性作用不明显(P>0.05)。荧光定量PCR结果显示,PGZ能分别下调和上调MMP-7 mRNA、TIMP-1 mRNA的表达,但TIMP-1 mRNA在实验组和对照组之间的差异无统计学意义。Matrigel侵袭实验和过河实验显示,随着PGZ浓度的升高,透过Matrigel膜的QBC939细胞数减少、过河时间延长,有剂量-效应关系(P<0.01)。结论PPAR-γ经其配体PGZ激活后能抑制QBC939细胞的体外侵袭力,其机制可能与调控MMP-7的表达及抑制细胞运动有关。

Objective To explore the effect and mechanism of ligand ploglitazone （PGZ） activating PPAR-γ on the invaslveness of cholanglocarclnoma cell in vitro. Methods Human hilar cholangiocarclnoma cell line QBC939 was selected and cultured in vitro for this research. The rate of QBC939 cell growth inhibition was detected by MTT, and the influence of PGZ on the expression of MMP-7 mRNA and TIMP-1 mRNA was measured by using FQ-PCR. The in vitro invastveness and mobility of QBC939 were quantified by Matrigel invasion assay and crossing-river test. Results Among the low concentration （5 μmol/L-40 μmol/L） groups at the point of 12-hours, PGZ did not show to inhibit significantly the growth of QBC939 cells （P〈0. 05）. However, the PGZ could down-regulate the expression of MMP-7 mRNA in QBC939 cells （P〈0.01）, and up-regulate the TIMP-1 mRNA expression to be without obvious statistics significance （P〉0. 05）. At last, PGZ could reduce the number of QBC939 cell breaking through the matrigel and prolonging the time of crosslng-river significantly （P〈 0.01） in a dose-dependent manner. Conclusion For ligand PGZ to activate PPAR-7 can inhibit the invasiveness of QBC939 cells in vitro via regulating the expression of MMP-7 and the mobility of QBC939 cells probably. [Key words]