摘要
目的探讨表皮生长因子通过核转录因子(NF)-кB诱导基质金属蛋白酶促进胆管癌细胞侵袭的机制。方法检测胆管癌细胞株HuCCT1在表皮生长因子(EGF)同浓度下的细胞侵袭力及细胞增殖情况；采用逆转录-聚合酶链反应(RT-PCR)及Western blot分析方法,检测基质金属蛋白酶(MMP)-2及MMP-9在不同EGF浓度下的基因和蛋白表达；采用凝胶电泳迁移率试验(EMSA)检测NF-кB在不同EGF浓度下的活性。结果随着EGF浓度的增加,胆管癌细胞株HuCCT1侵袭细胞数也随之增加,具有明显的浓度依赖性。而EGF浓度的变化对HuCCT1细胞的增殖无明显影响。EGF明显上调HuCCT1细胞中MMP-9的mRNA和蛋白表达水平,MMP-2不受影响。EGF增强NF-кB的活性；PDTC和布洛芬明显抑制EGF诱导的HuCCT1细胞侵袭力。结论EGF能够增强胆管癌细胞的侵袭力,其增强的侵袭力可能是通过NF-кB信号传导路径,激活MMP-9而实现的。
Objective To explore the mechanism by which epidermal growth factor （EGF） promotes human eholangioeareinoma cells invasion through NF-κB activating proteinase. Methods Invasion assay and eeU proliferation assay were performed in human eholangioeareinoma eeU lines HuCCT1 treated with different concentrations of EGF. RT-PCR and Western blot analysis were used to detect the expression of MMP-2, MMP-9 in human eholangioeareinoma cell lines HuCCT1. NF-κB activity was determined by eleetrophoretie mobility shift assay （EMSA）. In order to detect the effects of NF-κB on invasion of tumor cells, the cells were pretreated with pyrrolidine dithioearbamate （PDTC） or ibuprofen. Results Treatment with EGF resulted in an increased invasion of human eholangioeareinoma eeU lines in a dosedependent manner. However,EGF did not affect the proliferation in HuCCT1 cells. The results of RT- PCR and Western blot revealed that EGF dramatieaUy increased MMP-9, mRNA and protein expression in HuCCT1 eeUs, but that of MMP-2 was not changed. NF-κB activities were significantly increased by treatment with EGF, and EGF-indueed invasion of the HuCCT1 cells was inhibited by pretreatment with PDTC or ibuprofen. Conclusion EGF promotes the invasion of human eholangiocareinoma cells, and the up-regulation of MMP-9 and NF-κB activity may account for this invasion.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2006年第9期1092-1094,共3页
Chinese Journal of Experimental Surgery
关键词
表皮生长因子
基质金属蛋白酶
核转录因子-ΚB
胆管癌
侵袭
Epidermal growth factor
Matrix metalloproteinases
Nuclear factor-κB
Cholangioearcinoma
Invasion
