摘要
为了探讨半乳糖凝集素-7(Galectin-7)在食管癌Eca-109细胞发生发展中的细胞外功能及其机制,该实验采用免疫印迹法检测浓缩细胞上清液中Galectin-7的表达,并利用不同浓度的重组Galectin-7培养食管癌Eca-109细胞。使用免疫印迹、实时荧光定量PCR、细胞划痕实验、MTT实验检测在培养后细胞中基质金属酶-9(MMP-9)、p38、pp38表达水平的变化以及Eca-109细胞迁移和增殖能力的改变。结果显示,Galectin-7存在于Eca-109细胞细胞上清液中,加入重组Galectin-7培养细胞后,MMP-9、pp38的表达水平明显上升,并且细胞的迁移能力也得到了提高,增殖能力无明显变化。由此说明,在食管癌Eca-109细胞中,Galectin-7可以被细胞分泌至细胞外,可能通过结合细胞外膜上特异性糖基配体激活p38 MAPK通路诱导MMP-9的表达,从而在Eca-109细胞侵袭、迁移过程中发挥重要的作用。
To explore the extracellular function and mechanism ofGalectin-7 in the development of esopha- geal Eca-109 cell line, Western blot was used to detect the expression of Galectin-7 in the concentrated cell culture supernatant, and Eca-109 cell line were incubated with human recombinant Galectin-7 in different concentrations. Western blot, Real-tirne quantitative PCR, scratch test and MTT were used to analyze the expressions of MMP-9, p38 and pp38 in Eca-109 cell and the impact of Galectin-7 on the invasion and the proliferation of Eca-109 cell. The result testified the existence of Galectin-7 in the cell culture supematant, and the expressions of MMP-9 and pp38 were pro- moted by Galectin-7 and the ability of invading of Eca-109 was improved. Taken together, our results suggested that in Eca-109 cell line, Galectin-7 was secreted to the extracellular, and induced the expression of MMP-9 through activat- ing the p38 MAPK pathway by binding the specificity glycosylated receptor in the cell membrane.
出处
《中国细胞生物学学报》
CAS
CSCD
北大核心
2014年第12期1609-1615,共7页
Chinese Journal of Cell Biology