整合素β_1亚单位对肝癌细胞与层黏连蛋白趋化行为的影响

Integrin betal mediates hepatocellular carcinoma cells chemotaxis to laminin

目的 研究整合素β1亚单位对肝癌细胞与层黏连蛋白(LN)趋化行为的影响。 方法 采用双微吸管实验法进行肝细胞癌(HCC)细胞趋化实验,在两侧微管内加入相同浓度(50μg/ml,200 μg/ml)的LN,并引导微管尖端与同一HCC细胞紧密接触,动态观察细胞两侧伪足形成过程;当两侧微管LN浓度为200μg/ml,在一侧微吸管内加入针对整合β1亚单位(CD29)的单克隆抗体(Anti-CD29)20μg/ml,考察β1整合素亚单位阻断对HCC细胞伪足形成的影响。利用流式细胞仪对HCC细胞表面整合素β1亚单位的表达进行分析。 结果 两侧微吸管加入相同浓度的LN,细胞向两侧微吸管两侧均有伪足形成;随作微吸管内LN浓度的增加,细胞伪足增加,且两侧微吸管内伪足的变化基本呈对称性;在此基础上,微吸管加入Anti-CD29的一侧,HCC细胞伪足生长曲线呈现明显的抑制,而未加入Anti-CD29的微吸管一侧与加入的对侧相比,该侧细胞的伪足明显增加。流式细胞仪分析表明,所研究细胞整合素β1亚单位表达率达95.78％。 结论 整合素β1亚单位是联系HCC细胞与HCC细胞对LN趋化性伪足形成的重要受体基础。

Objectives To study the effects of integrin betal on the chemotaxis of hepatocellular carcinoma (HCC) cells to laminin (LN). Methods A micropipette technique was adopted to investigate the effect of integrin betal blockade on pseudopod protrusion of HCC cells in response to LN stimulation. Chemotactic pseudopod protrusion of a HCC cell was evaluated using a dual-pipette set-up, in which two pipettes filled with LN solution were positional in close contact with the same cell, and pseudopod protrusion into each pipette was viewed dynamically and recorded with a tape recorder. The lengths of pseudopods were measured, then plotted against time to obtain a pseudopod growth curve. The integrin betal subunit on the surfaces of HCC cells was analyzed by flow cytometry. Results In dual pipette chemotaxis experiment, when the two pipettes were filled with LN (50 μg/ml, 200μg/ml), pseudopods extended from the HCC cells into each of the pipettes nearly symmetrically. Upon addition of anti-CD29 (20 μg/ml) to one of the pipettes, the pseudopod protrusion was blocked almost completely, while the pseudopod protrusion into the opposite pipette became more evidently, with larger maximum length. The expression rate of integrin betal on the cells was up to 95.78% Conclusion Integrin betal subunit is the important receptor for mediating HCC cells chemotaxis to laminin.