甲基莲心碱抑制K562/A02细胞GSTπ的表达
白血病,,白血病;多药耐药;甲基莲心碱;谷胱甘肽S转移酶π,1材料与方法,2结果,3讨论,参考文献
摘要:目的 探讨甲基莲心碱(Neferine, Nef)对谷胱甘肽S转移酶π(GSTπ)的影响。 方法 应用MTT法测定甲基莲心碱处理的K562/A02细胞对阿霉素(ADM)敏感性的影响。RTPCR法分析Nef处理后,GSTπmRNA转录的变化。通过Western Blot观察Nef对GSTπ蛋白表达的影响。结果 甲基莲心碱对K562/A02的耐药性具有明显的逆转作用。用Nef处理后,对阿霉素药物敏感性的相对逆转效率第1天、第3天和第5天分别为9.6%、41.4%和10.7%;GSTπmRNA转录水平在用药后第3天下调最大,为(50.7±2.3)%。GSTπ蛋白的表达于用药后第1天、第3天和第5天分别下降25.57%、48.42%和10.68%。结论 甲基莲心碱具有较强的逆转K562/A02细胞多药耐药的作用,其逆转机制可能包括下调K562/A02细胞GSTπmRNA的转录和蛋白的表达。关键词:白血病;多药耐药;甲基莲心碱;谷胱甘肽S转移酶π
Effects of neferine on expression of glutathione
Stransferasepi in K562/A02 cells in vitroXiao Xibin1, Xie Zhaoxia1, Qin Qun2
(1. Department of Hematology; 2. Department of Pharmacy, Xiangya Hospital,Central South University, Changsha 410008, China)ABSTRACT: Objective To study the effects of neferine on the expression of glutathione Stransferasepi in vitro and to explore the multidrug resistance reversing mechanism of neferine. Methods 50% inhibition concentration (IC50) of ADM on K562/A02 was determined by MTT method. The transcription of GSTπ gene was detected by semiquantitative RTPCR and the expression level of GSTπ was determined by Western blot after neferine treatment. Results Neferine remarkably enhanced chemosensitivity to ADM of K562/A02 cells. After neferine treatment in day 1, day 3 and day 5, the relative efficiency of K562/A02 to ADM was 9.6%, 41.4% and 10.7%, respectively. Semiquantitative RTPCR showed that mRNA transcription of GSTπ gene was significantly reduced (P<0.01).Neferine was shown to efficiently inhibit the expression of GSTπ according to the results of Western blot (P<0.05). Conclusion Increase of GSTπ expression is one of the MDR mechanisms in K562/A02 cells. Neferine could modulate GSTπ expression in a timedependent manner. Decrease of GSTπ expression may be one of the MDR reversal mechanisms in K562/A02 cells by neferine. ......
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