D-丙2,D-亮5脑啡肽对人肝癌细胞生长、迁移能力的影响及机制研究

Effect of [D-Ala2, D-Leu5] encephalin(DADLE) on cell migration and growth in human hepatocellular carcinoma and its mechanisms

目的:探讨δ阿片受体激动剂D-丙2,D-亮5脑啡肽(DADLE)对人肝癌细胞生长及迁移能力的影响及机制。方法:分别以不同浓度的DADLE(0.01、0.1、1.0和10μmol/L)处理对数生长期的HepG2细胞,MTT法检测细胞增殖活力的变化,划痕实验观察细胞迁徙能力的变化,RT-PCR和Western blot法检测PKC(蛋白激酶C)、IQGAP1(含IQ结构的GTP酶活化蛋白1)的mRNA和蛋白表达变化。结果:与对照组相比,加入不同浓度的DADLE后,MTT法显示肝癌细胞的增殖能力明显增加,并且具有浓度依赖性。划痕实验显示加入DADLE可明显增强肝癌细胞的迁徙能力;RT-PCR和Western blot显示加入DADLE可显著增加肝癌细胞内PKC和IQGAP1的mRNA和蛋白表达水平。结论:DADLE激活δ阿片受体对人肝癌细胞的生长及迁徙具有促进作用,其机制可能与增加PKC和IQGAP1表达实现的。

Objective To study[D-Ala2, D-Leu5] enkephalin （DADLE）＇s effects on cell migration and growth in human hepatocellular carcinoma （HCC） and its mechanisms. Methods HepG2 cells were treated with different concentration of DADLE （0.01, 0.1, 1.0 and 10 μmol/L）. MTY assay was used to detect the cell proliferation. Scratch adhesion test was used to observed cell migration ability. RT-PCR and Western blot was used to detect the expression levels of PKC and IQGAP1. Results The proliferation of HCC cells was increased significantly after treatment with different concentration of DADLE and this effect was a concentration-dependent manner. The results of scratch adhesion test showed that HCC cells migration ability were obviously encreased. The results of RT-PCR and Western blot showed that DADLE encreased the PKC and IQGAPlexpression in HCC cells. Conclusion DADLE encreased HCC cells migration and growth through activating delta opioid receptor, which could be associated with increasing the PKC and IQGAPlexpression.