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目的:用三维培养方法在体外模拟体内实体瘤生长情况,探讨结肠癌细胞群集耐药与细胞黏附分子整合素αⅤ(integrin αⅤ)和整合素β3(integrin β3)过度表达的关系。方法:运用三维培养法获得人结肠癌细胞HT29多细胞球体(multicellular spheroids,MCS)模型,采用RT-PCR法及免疫荧光法比较单层细胞(monolayer cells,MC)及MCS细胞中基质间黏附分子integrin αⅤ和integrin β3 mRNA及蛋白表达水平变化,用FCM法检测5-氟尿嘧啶(5-fluorouracil,5-FU)作用48h后细胞的生存率和凋亡率。结果:HT29 MCS与HT29 MC经比较,integrin αⅤ和integrin β3 mRNA及蛋白表达水平均明显增高(P<0.01),对5-FU的药物敏感性显著下降(P<0.05),5-FU诱导的细胞凋亡率显著下降(0.346±0.035vs0.235±0.024),差异均有统计学意义(P<0.05)。结论:细胞黏附分子integrin αⅤ和integrin β3表达水平增高可能增加结肠癌细胞群集的耐药性。
OBJECTIVE: To study the growth of solid tumors in vivo by three-dimensional culture in vitro and to explore the relationship between colon cancer cell cluster resistance and the overexpression of integrin αⅤ and integrin β3. Methods: The human colon cancer cell HT29 multicellular spheroids (MCS) model was obtained by using three-dimensional culture method. The expression of intercellular adhesion molecule (MMP) in monolayer cells (MC) and MCS cells was analyzed by RT-PCR and immunofluorescence The changes of integrin αⅤ and integrin β3 mRNA and protein levels were detected by FCM. After 5-fluorouracil (5-FU) treatment for 48h, the cell viability and apoptosis rate were detected. Results: Compared with HT29 MC, the mRNA and protein expressions of integrin αⅤ and integrin β3 were significantly increased (P <0.01) and the drug sensitivity to 5-FU was significantly decreased (P <0.05) The apoptosis rate was significantly decreased (0.346 ± 0.035 vs 0.235 ± 0.024), the differences were statistically significant (P <0.05). Conclusion: Increased expression of integrin αⅤ and integrin β3 may increase the drug resistance of colon cancer cells.