硫酸右旋糖苷抑制人胃癌细胞黏附以及整合素β1蛋白表达机制研.docVIP

硫酸右旋糖苷抑制人胃癌细胞黏附以及整合素β1蛋白表达机制研 　 作者：徐远义，黄允宁，王炜，马竟先，董俭达，柳勇，高宏，曹相枚，赵琳 【摘要】 ［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。 【关键词】 胃肿瘤 The Mechanism of Dextran Sulfate Inhibiting Cells’ Adhesion and Integrin β1 Expression in Human Gastric Cancer Cell Line Abstract: ［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 immunofluorescent cytochemistry 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 cells 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. MKN1 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% (110kDa) 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. Key words: dextran sulfate; gastric neoplasms; integrin β1 胃癌腹膜种植转移的机制尚不十分清楚。一般认为癌细胞浸润胃壁浆膜层并脱离癌组织母体进入腹腔，成为有生物活性的脱落癌细胞，黏附于腹膜并增殖形成种植转移灶[1]。在这过程中细胞黏附是一个