水通道蛋白9在结肠癌组织中的表达变化及其过表达对结肠癌细胞侵袭和迁移的影响

Expression changes of aquaporin 9 in colon cancer tissues and its effect on invasion and migration of colon cancer cells

目的观察结肠癌组织中水通道蛋白9(AQP9)的表达情况,分析结肠癌HCT116细胞中AQP9过表达对肿瘤细胞侵袭、迁移能力的影响。方法收集33例结肠癌患者的结肠癌组织及癌旁正常组织,采用免疫组化IHC法检测AQP9表达。取结肠癌HCT116细胞,分为空白组、空质粒组和AQP9过表达组,空质粒组和AQP9过表达组分别转染pcDNA3.1和AQP9过表达质粒,空白组为未经处理的细胞。采用Transwell侵袭实验观察三组细胞侵袭能力,细胞划痕实验观察24 h细胞迁移的剩余面积来评价细胞迁移能力,Western blotting法检测细胞上皮间充质转化(EMT)相关蛋白E-cadherin、Vimentin的相对表达量。结果AQP9在结肠癌组织中的阳性表达率低于癌旁正常组织(P<0.05)。AQP9过表达组穿膜细胞数较空白组、空质粒组减少(P均<0.05),24 h细胞迁移剩余面积较空白组、空质粒组减小(P均<0.05)。AQP9过表达组E-cadherin表达较空白组、空质粒组升高,Vimentin表达较空白组、空质粒组降低(P均<0.05)。结论结肠癌组织中AQP9表达降低;AQP9过表达可抑制结肠癌的侵袭和迁移能力,其机制可能是通过抑制EMT来实现的。

Objective To observe the expression of aquaporin 9(AQP9)in the colon cancer tissues and to analyze the effect of overexpression of AQP9 on invasion and migration of colon cancer HCT116 cells.Methods Colon cancer tissues and adjacent normal tissues of 33 patients with colon cancer were collected.The expression of AQP9 was detected by immunohistochemical IHC method.HCT116 cells were divided into the blank group,blank plasmid group,and AQP9 overexpression group.HCT116 cells in the blank plasmid group and AQP9 overexpression group were transfected with pcDNA 3.1 and AQP9 overexpression plasmids,respectively;the blank group was untreated.Transwell invasion test was used to observe the invasive abilities of three groups.We used cell scratch assay to observe the remaining area of 24 hour cell migration to assess the migration abilities.Western blotting was used to detect the relative expression of E-cadherin and Vimentin related proteins in epithelial mesenchymal transformation(EMT).Results The positive expression rate of AQP9 in the colon cancer tissues was lower than that in the adjacent normal tissues(P<0.05).The number of transmembrane cells in the AQP9 overexpression group was lower than that in the blank group and the empty plasmid group(P<0.05),and 24 h cell migration remaining area was smaller than that in the blank group and blank plasmid group(P<0.05).The expression of E-cadherin in the AQP9 overexpression group was higher than that in the blank group and blank plasmid group,while the expression of Vimentin was lower(both P<0.05).Conclusion The expression of AQP9 decreases in colon cancer tissues,and the overexpression of AQP9 can inhibit the invasion and migration of colon cancer,which may be achieved by inhibiting EMT.