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目的通过观察人胃癌MKN1细胞的粘附过程和测定整合素β1cDNA的表达水平,研究硫酸右旋糖苷(dextransulfate,DS)对人胃癌MKN1细胞粘附的抑制作用的机制。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和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RT-PCR测定整合素β1的cDNA表达。结果MKN1细胞通过变形,形成伪足粘附到培养盘,并与其他细胞接触形成单细胞层。免疫染色可见整合素β1在细胞膜上有强烈的表达,并且形成整合素集落。DS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;使MKN1细胞的伪足缩短,表现出维持圆形的倾向。在使用DS培养4h后仍有部分细胞处于游离状态,分别收集游离细胞和贴璧细胞,用RT-PCR测定整合素β1的cDNA表达。实验组贴壁细胞和游离细胞内整合素β1的cDNA的表达水平分别减少到对照组细胞的74%和38%。结论DS对人胃癌细胞MKN1粘附过程的抑制与整合素β1表达的减弱有关。
OBJECTIVE: To investigate the mechanism of dextransulfate (DS) inhibition on the adhesion of human gastric cancer MKN1 cells by observing the adhesion process of human gastric cancer MKN1 cells and measuring the expression of integrin β1 cDNA. Methods MKN1 cells were cultured with DS or PBS, respectively. The adherent process and morphological changes of fixed cells and living cells on the culture plate were observed by fluorescence immunocytochemical staining and confocal microscopy. The integrin β1 cDNA expression was measured. As a result, MKN1 cells adhered to the culture plate by deforming to form pseudopods, and contacted with other cells to form a monolayer. Immunostaining showed that integrin β1 was strongly expressed in the cell membrane and formed integrin colonies. DS inhibits the expression of integrin β1 and decreases the area of integrin colonies that have formed; pseudopodia of MKN1 cells are shortened, showing a tendency to remain circular. After cultured with DS for 4h, some cells remained free and free cells and adherent cells were collected respectively. The expression of integrin β1 was determined by RT-PCR. The expression of integrin β1 in the adherent cells and free cells in the experimental group decreased to 74% and 38% of the control cells, respectively. Conclusion DS inhibits the adhesion of human gastric cancer cell line MKN1 to the decrease of integrin β1 expression.