摘要
目的:研究乏氧对人食管鳞状细胞癌EC9706细胞株的多药耐药相关蛋白(MRP)表达的影响,探讨乏氧诱导因子-1α(HIF-1α)在乏氧引起肿瘤细胞多药耐药中的作用。方法:采用化学性低氧诱导剂氯化钴(CoCl2)体外培养人食管鳞状细胞癌EC9706细胞株,用RNAi沉默HIF-1α基因表达,应用半定量RT-PCR检测HIF-1α基因沉默效果。应用RT-PCR、免疫细胞化学技术检测RNA干扰前后乏氧条件下MRP1蛋白及RNA水平表达的变化。结果:乏氧后EC9706细胞中MRP1 mRNA和蛋白的表达均增加(P<0.05)。针对HIF-1α基因的siRNA干扰技术,能有效沉默HIF-1α基因的表达。同样乏氧条件,转染HIF-1α siRNA后EC9706细胞较未转染及转染Control siRNA组细胞相比,MRP1 mRNA和蛋白的表达均减少(P<0.05)。结论:HIF-1α上调食管鳞状细胞癌细胞中MRP1的表达可能是乏氧条件下化疗耐药的机制之一;HIF-1α可望成为新的食管鳞状细胞癌治疗的靶点。
Aim ： To study the effect of hypoxia on expression of multidrug resistance protein（MRP1 ） and its mechanism of muhidrug resistance in ESCC, and to provide a new target for treatment of ESCC. MethOdS ：To establish a hypoxia model of EC9706 cells by a chemical hypoxia inducer （ Cobalt chloride, CoC12 ）. RNA-interference targeting HIF-1 α was used to silence the expression of HIF-1α in human EC9706 cell line. The effect of HIF-1α gene silencing was measured by RT-PCR. The expressions of MRP1 were measured respectively by RT-PCR, immunocytochemistry in EC9706 cells under normia, hypoxia and after RNA interference under hypoxia. Results： HIF-1α mRNA expression was obviously knocked down by small interferencing RNA（siRNA）. Expression of MRP1 on mRNA and protein level was significantly increased in EC9706 cells after hypoxia（ P 〈 0.05 ）. MRP1 expression on mRNA and protein level in cells transfected with HIF-1 α siRNA was significantly lower than those in untransfeeted group and transfeeted with control siRNA group under hypoxia（P 〈 0.05）. Conclusion ： HIF-1α can upregulate the expression of MRP1, which may involve in the chemotherapeutic resistance of EC9706 cells under hypoxia. HIF-1α can be used as a new, tumor-specific target for anticancer therapy in ESCC.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2009年第1期51-55,共5页
Journal of Zhengzhou University(Medical Sciences)
