人小细胞肺癌细胞H446Snail基因表达及其在顺铂耐药中的作用.docVIP

人小细胞肺癌细胞H446 Snail基因表达及其在顺铂耐药中的作用 况里杉, 周向东 (400038重庆,第三军医大学西南医院呼吸内科) [摘要]　目的 观察锌指转录因子snail在耐顺铂（cisplatin,CDDP）的人小细胞肺癌(small cell lung cancer, SCLC)H446/CDDP细胞中的表达情况，探讨snail表达在人SCLC顺铂耐药中的作用。 方法　 培养H446细胞系和H446/CDDP细胞系，采用MTT法检测H446/CDDP的耐药指数；分别提取人SCLC顺铂耐药细胞H446/CDDP及其亲代细胞H446的mRNA及总蛋白，应用sybgreen实时荧光RT-PCR、western-blot方法检测snail mRNA及蛋白表达水平。结果　H446/CDDP细胞snail蛋白及mRNA表达水平较H446细胞显著性升高role in cisplatin resistance Kuang Lisha 1, Zhou Xiangdong 1 (1Department of Respiratory Diseases, southwest Hospital, Third Military Medical University, Chongqing, 400038, China) [Abstract]　Objective　we observed the expression of zinc finger transcription factor Snail in the human cisplatin resistance small-cell lung cancer(SCLC) cell line H446/CDDP, so as to study its role in human SCLC cisplatin resistance . Methods　H446 and H446/CDDP cell lines were Cultured, cisplatin resistance indexes were measured by methyl tetrazolium(MTT) assay. mRNA and total protein are respectively extracted from H446 and H446/CDDP.Expression level of snail in mRNA and protein level was detected by real time RT-PCR and western-blot. Results　The snail expression of H446/CDDP cells were significantly higher than H446 cells both in protein and mRNA level. Conclusion The ciaplatin resistance in human SCLC may be relevant to the high expression level of snail . [Key words]　human small cell lung cancer; cisplatin resistance; snail；real time RT-PCR；western-blot Supported by the National Basic Research Program of China ( 973 Program，2010CB529402) ． Corresponding author: Zhou Xiangdong, E-mail: xiangdongzhou@126.comSCLC 易发生顺铂耐药，其机制目前尚不完全清楚，可能与凋亡、DNA修复异常、细胞内药物异常积累等多种因素相关[1]。Snail(锌指转录因子)是一种抑制型转录因子，通过直接抑制钙黏蛋白E(E-cadherin) 上皮-间充质转化(epithelial to mesenchymal transition,EMT)[2, 3]。在肺腺癌细胞系A549的研究中发现沉默Snail的表达能够恢复细胞对顺铂的敏感性[4, 5]，而在SCLC尚缺乏相关方面的研究。因此，本研究拟从EMT的关键因子snail入手,研究人SCLC顺铂耐药性的获得与snail表达水平之间的关系,为逆转其耐药寻求新的途径。 [基金项目]　国家重点基础研究发展计划（973计划，2010CB529402） [通信作者]　周向东, E-mai: xiangdongzho