摘要
目的观察沉默孕烷X受体(pregnane X receptor,PXR)表达对结肠癌细胞奥沙利铂化疗敏感性的影响,初步探讨其可能机制。方法构建PXR基因的siRNA质粒表达载体,转染结肠癌LS17T细胞,并建立稳定表达细胞,用RT-PCR和Western blot方法检测特异性蛋白1(specificity protein 1,SP1)和多药耐药相关蛋白3(multidrug resistance-asso-ciate protein 3,MRP3)表达,MTT方法检测奥沙利铂化疗敏感性。结果成功建立PXR敲低细胞克隆PXRi 1#、PXRi 2#;在PXRi 1#和PXRi 2#细胞克隆中,MRP3 mRNA及蛋白表达明显下降,SP1表达也下降;与PXRi control相比,细胞克隆PXRi 1#、PXRi 2#奥沙利铂的IC50值均显著降低[(4.83±0.22)、(4.75±0.19)vs(9.44±0.37),P<0.05]。结论 PXR表达沉默可增强结肠癌LS174T细胞对奥沙利铂化疗敏感性,这可能与PXR沉默后,SP1、MRP3表达下调相关。
Objective To determine the effect of silencing pregnane X receptor （PXR） expression by RNA interference on the chemosensitivity of LS174T cells to oxaliplatin. Methods The PXR RNAi expression plasmids were constructed, and then transfected LS174T cells to establish stable transfection cell clones. The expression of specificity protein （SP） 1 and muhidrug resistance-associate protein 3 （MRF3） were detected by RT-PCR and Western blotting. The chemosensitivity of the transfected and nontransfected cells to oxaliplatin was evaluated by MTI＂ assay. Results The cell clones PXRi 1 # and PXRi 2# with stable PXR silence were constructed successfully. In cell clones PXRi 1# and PXRi 2#, the expression of MRP3 at mRNA and protein levels was decreased markedly, and that of SP1 also was decreased. Compared with control group, the ICs0 of PXRi 1# and PXRi 2# cells to oxaliplatin was significantly decreased （4.83 ＋0.22 and 4.75 _＋0. 19 vs 9.44 _＋ 0.37, P 〈 0.05 ）. Conclusion Interference of PXR expression can elevate the chemosensitivity of LS174T cells to oxaliplatin, which might be through PXR knock-down downregulating SP1 and MRP3.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2012年第1期5-8,共4页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(81172114)
四川省卫生厅科研课题(100188)~~
关键词
孕烷X受体
结肠肿瘤
多药耐药
奥沙利铂
pregnane X receptor colon neoplasms multidrug resistance oxaliplatinSupported by the General Program of National Natural Science Foundation of China （ 81172114 ） and the Project of Medical Research of Sichuang Health Depart-ment （100188）. Corresponding author： Liang Houjie, Tel： 86-23-68754128, E-mail： lianghoujie@ sina. com
