结肠癌HT29细胞的相关分子表达及与多药耐药的相关性

目的:探讨结肠癌HT29细胞的相关分子表达及与多药耐药的相关性。方法:选择人结肠癌HT29细胞株进行三维培养，采用RT-PCR与免疫组化检测整合素ανβ的表达情况，同时在三维培养和整合素ανβ3阻断型抗体加入条件下对细胞的多药化疗耐药性进行检测。结果:结肠癌HT29细胞在三维培养24h后，部分聚集成由数个细胞组成的细胞团，电镜观察多细胞球球体呈较规则的致密的球形结构，多细胞球球体模型构建成功。结肠癌HT29细胞多细胞球球体中整合素ανβ3基因的表达比一般平面培养细胞高，整合素β3蛋白、整合素αν主要定位于胞浆，在多细胞球球体中表达明显高于普通平面培养细胞。HT29细胞多细胞球球体+整合素ανβ3抗体组在5-氟尿嘧啶和奥沙利铂作用后48h的细胞凋亡率为0.31±0.04，细胞坏死率为0.52±0.02；而HT29细胞多细胞球球体组的细胞凋亡率与坏死率分别为0.14±0.03和0.21±0.03，对比差异明显(P<0.05)。结论:结肠癌HT29多细胞球球体中的整合素ανβ3在基因与蛋白水平都呈现表达升高的趋势，可能是细胞多药耐药产生的重要原因。

The related molecules expression and multidrug resistance correlation in the HT29 colon cancer cell

Objective:To investigate the related molecules expression and multidrug resistance in HT29 colon cancer cell.Methods:To choose HT29 human colon cancer cell line for the three-dimensional culture，RT-PCR and immunohistochemistry were used for detecting the expression of integrin ανβ.Results:HT29 colon cancer cells in three-dimensional culture after 24h were showed gather consists of several cells，and electron microscopy showed the multicellular balls substantially more regular dense spherical structure that multicellular spheres model were successfully constructed.ανβ3 integrin gene expression in colon cancer HT29 cells in multicellular spheres were significantly higher than conventional flat cultured cells.The integrins αν，integrin β3 protein were mainly localized in the cytoplasm，expressed in multicellular spheres were significantly higher than normal plane cultured cells.The after 5-fluorouracil and oxaliplatin for 48h apoptosis and cell necrosis in the HT29 cells multicellular spheres+integrin antibody group ανβ3 group were 0.31±0.04 and 0.52±0.02，so that were 0.14±0.03 and 0.21±0.03 that compared significantly different（P<0.05).Conclusion:Colon cancer HT29 multicellular spheres of integrin ανβ3 in gene expression and protein levels has increasing trend that may be an important reason of multidrug resistance generated.