硫酸右旋糖苷抑制人胃癌细胞黏附以及整合素β_1蛋白表达的机制研究

The Mechanism of Dextran Sulfate Inhibiting Cells' Adhesion and Integrin β_1 Expression in Human Gastric Cancer Cell Line

[目的]探讨硫酸右旋糖苷(DS)对人胃癌MKN1细胞的黏附抑制作用机制。[方法]分别在培养液中加入DS或PBS对MKN1细胞进行培养,用荧光免疫细胞染色法进行染色,用共聚焦显微镜对固定细胞和活细胞在培养盘上的贴壁过程及形态变化进行观察,用Western blotting对整合素β1蛋白的表达水平进行测定。[结果]MKN1细胞形成伪足并与其他细胞接触形成单细胞层。整合素β1在细胞膜上有强表达并形成集落。DS使MKN1细胞的伪足缩短,表现出维持圆形形状的倾向;抑制整合素β1的表达并使已经形成的整合素集落区域减少。MKN1细胞表达了整合素β1的两种形式(130kDa和110kDa),实验组贴壁细胞整合素β1蛋白的表达分别减少到对照组细胞的64%(130kDa)和57%(110kDa),实验组游离细胞整合素β1蛋白的表达分别减少到对照组细胞的51%(130kDa)和15%(110kDa)。[结论]DS对人胃癌细胞MKN1黏附过程的抑制与整合素β1蛋白表达的减少有关。

[Purpose] To explore mechanism of the adhesion of human gastric cancer MKN1 cells inhibited by destran sulfate （DS）. [Methods] MKN1 cells were cultured with DS or PBS, then stained with immunofluoreseent eytoehemistry and observed in fixed or living conditions by confocal laser scanning microscope. Western blot was used to analyze the protein expression of MKN1 cells. [Results] MKN1 cells formed filopodia and adhered to other eells to form a cell-monolayer. Integrin β1 intensively expressed in the cell membrane and formed clusters. DS-treated cells tended to maintain a round shape by contracting the filopodia. DS inhibited the expression of integrin β1 and decreased the area of integrin β1 clusters. MKNI cells expressed two forms of integrin β1（130kDa and 110kDa）. Both forms of integrin β1 protein expression of the attaching cells in the experimental group decreased to 64%（130kDa） and 57% （l10kDa） of those in the control group, the integrin β1 protein expression of the floating cells in the experimental group decreased to 51%（130kDa） and 15%（110kDa） of that in the control group. [Conclusion] DS inhibiting MKN1 cells＇ adhesion is related to decrease of integrin β1 expression.