摘要
目的:研究可溶性耐药相关钙结合蛋白(sorcin)在人胶质瘤细胞对顺铂敏感性中的作用。方法:构建pSilencerTM3.1-H1-sorcin siRNA表达质粒;质粒转染人胶质瘤U251细胞;RT-PCR和Western blotting法检测sorcin mRNA和蛋白表达的变化;MTT法检测U251细胞的生存率;Western blotting检测耐药相关蛋白的表达变化。结果:经酶切和测序鉴定,成功构建pSilencerTM3.1-H1-sorcin siRNA表达载体;将该表达载体转染U251细胞,RT-PCR和Western blotting结果显示sorcin mRNA和蛋白表达量降低(P<0.05);MTT结果显示,抑制sorcin表达可增强U251细胞对顺铂的敏感性(P<0.05);同时发现抑制U251细胞的sorcin表达能降低耐药相关蛋白P-糖蛋白(P-gp)和多药耐药相关蛋白1(MRP1)的蛋白水平(P<0.05)。结论:抑制sorcin表达增强U251细胞对顺铂的敏感性,其作用机制可能与降低耐药相关蛋白P-gp和MRP1的表达有关,提示sorcin可能与胶质瘤细胞顺铂耐药有关。
AIM： To investigate the effect of sorcin expression on the sensitivity of human glioma cells to cisplatin.METHODS： pSilencerTM3.1-H1-sorcin siRNA recombinant plasmid was constructed,and transfected into human glioma U251 cells.RT-PCR and Western blotting were used to analyze the expression of sorcin at mRNA and protein levels after transfection.The viability of U251 cells was measured by MTT assay.The protein expression of P-glycoprotein（P-gp） and multidrug resistance-associated protein 1（MRP1） in U251 cells was detected by Western blotting.RESULTS： The plasmid pSilencerTM 3.1-H1-sorcin siRNA was successfully constructed,and was confirmed by restriction enzyme digestion and sequence analysis.The expression of sorcin at mRNA and protein levels was significantly decreased after sorcin siRNA was transfected into U251 cells（P 0.05）.Inhibition of sorcin expression significantly decreased the viability of U251 cells treated with cisplatin（P 0.05）,and the expression of P-gp and MRP1 proteins was also inhibited（P 0.05）.CONCLUSION： Inhibition of sorcin expression increases the sensitivity of U251 cells to cisplatin by decreasing the expression of resistance-related proteins P-gp and MRP1,suggesting that sorcin may be associated with the resistance of glioma cells to cisplatin.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第7期1202-1206,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81272876
No.81141099
No.81202552)
关键词
神经胶质瘤
顺铂
耐药性
可溶性耐药相关钙结合蛋白
Glioma
Cisplatin
Drug resistance
Soluble resistance-related calcium-binding protein
