Lewisy抗原对人卵巢癌细胞RMG-I-H部分耐药相关蛋白基因表达的影响

Effect of Lewis y Antigen on Regulating Gene Expression of Partial Drug Resistance Associated Proteins in Human Ovarian Cancer Cell Line RMG-I-H

目的探讨细胞表面Lewisy抗原对人卵巢癌细胞系RMG-I-H细胞的部分耐药相关蛋白基因表达的影响。方法RT-PCR法检测转染α1,2-岩藻糖转移酶基因和未转染α1,2-岩藻糖转移酶基因的人卵巢癌细胞系RMG-I-H和RMG-I细胞中,以及10μg/ml抗Lewisy单克隆抗体处理及未处理的RMG-I-H细胞中部分耐药相关蛋白mRNA的表达量。免疫细胞化学方法检测RMG-I和RMG-I-H细胞中P-糖蛋白(P-gp)的表达;分别建立RMG-I和RMG-I-H细胞的裸鼠皮下移植瘤模型,采取免疫组织化学方法检测移植瘤组织P-gp的表达。结果与RMG-I细胞比较,RMG-I-H细胞中蛋白激酶C-α(PKC-α)、拓扑异构酶Ⅰ(TopoⅠ)、多药耐药相关蛋白-1(MRP-1)及MRP-2基因mRNA的相对表达量均显著增高(0.46±0.02vs.0.27±0.05、0.82±0.08vs.0.52±0.04、0.66±0.07vs.0.34±0.12和0.44±0.08vs.0.23±0.05,均P<0.05),而多药耐药基因-1(MDR-1)mRNA的相对表达量显著降低(0.26±0.05vs.0.45±0.08,P<0.05)。免疫化学染色分析显示,不论体内还是体外,P-gp在RMG-I-H细胞中的表达量均显著高于RMG-I细胞中的表达量(均P<0.05)。经Lewisy单克隆抗体处理后,RMG-I-H细胞中MDR-1、MRP-1、MRP-2、PKC-α及TopoⅠ的mRNA相对表达量均呈时间依赖性下降(均P<0.05),而对照组以上基因mRNA的表达量不随时间的变化而变化(均P>0.05)。经Lewis y抗体处理6h的RMG-I-H细胞中MDR-1、MRP-1、MRP-2、PKC-α及TopoⅠ的mRNA相对表达量均显著低于对照组(均P<0.05),Lewis y单克隆抗体对上述基因表达的抑制率分别为48.55%、77.50%、70.18%、45.86%和46.13%。结论人卵巢癌细胞表面Lewisy抗原与部分耐药相关蛋白基因表达的调节密切相关。

Objective To investigate the influence of Lewis y antigen on the gene expression of partial drug resistance associated proteins in human ovarian cancer cell line RMG-I-H. Methods RT-PCR was used to determine the gene expressions of partial drug resistance associated proteins in RMG-I-H cell line transfected with α1,2-fucosyltransferases gene and RMG-I cell line,as well as in RMG-I-H treated with or without anti-Lewis y monoclonal antibody at the concentration of 10 μg/ml. The immunocytochemical method was used to detect the expression of P-glycoprotein （P-gp） in RMG-I and RMG-I-H cell lines. RMG-I and RMG-I-H cells were transplanted into nude mice and the expression of P-gp in the tissues was measured by immunohistochemistry. Results The mRNA expressions of protein kinase C-α（PKC-α）,topoismeraseⅠ（Topo Ⅰ）,multidrug resistance-associated protein-1（MRP-1）,and MRP-2 were significantly higher in RMG-I-H cells than those in RMG-I cells （0.46±0.02 vs. 0.27±0.05,0.82±0.08 vs. 0.52±0.04,0.66±0.07 vs. 0.34±0.12,and 0.44±0.08 vs. 0.23±0.05;all P〈0.05）. However,the mRNA expression of multidrug resistance 1 （MDR-1） was significantly lower in RMG-I-H cells than that in RMG-I cells （0.26±0.05 vs. 0.45±0.08,P〈0.05）. The P-gp level increased in RMG-I-H cells compared with that in RMG-I cells both in vivo and in vitro （P〈0.05）. Expressions of MDR-1,MRP-1,MRP-2,PKC-α,and TopoⅠ mRNA decreased by the time in RMG-I-H cells treated with anti-Lewis y monoclonal antibody （all P〈0.05）,while mRNA expressions of those genes in the control group did not statistically change （P〉0.05）. In addition,MDR-1,MRP-1,MRP-2,PKC-α,and TopoⅠ mRNA expressions were significantly lower in RMG-I-H cells treated with anti-Lewis y monoclonal antibody than those in the control group at 6 hours （all P〈0.05） and the inhibition ratios were 48.55%,77.50%,70.18%,45.86%,and 46.13%,respectively. Conclusion The Lewis y antigen of the human ovarian cancer cell surface is closely correlated with the regulation on the gene expression of partial drug resistance associated proteins.