【摘要】 本研究旨在探讨汉防己甲素(Tet)在逆转K562/A02细胞耐药与耐药相关的可溶性钙结合蛋白(SORCIN)基因表达之间的关系,为Tet的临床应用提供新的理论基础。通过MTT法筛选出所需浓度Tet,与K562/A02细胞株孵育培养48小时后,通过MTT检测Tet对柔红霉素(DNR)细胞毒性的影响,用RT-PCR方法检测SORCIN基因表达的变化,用Western blot方法检测sorcin表达蛋白的变化。结果表明:在所选浓度范围内,Tet可以增强DNR对K562/A02的细胞毒作用,(Tet浓度为0,0.5,1.0,2.0mg/L时DNR和Tet的IC50分别为11.3±0.17,5.15±0.10,3.91±0.06,2.52±0.04mg/L,p<0.01);K562/A02细胞中SORCIN基因高表达,且随着Tet浓度的增加SORCIN的表达先增强后减弱(Tet浓度为0.5mg/L时增强,1.0和2.0mg/L时减弱,p<0.05);K652细胞中sorcin蛋白低表达,K562/A02细胞中sorcin蛋白高表达,且随着Tet浓度的增加先增强后减弱(Tet浓度为0.5mg/L时增强、1.0和2.0mg/L时减弱,p<0.05)。结论:MTT检测显示Tet对K562/A02耐药逆转呈浓度依赖性,Tet可能通过调整SORCIN基因及蛋白的表达参与逆转K562/A02的耐药性,但并不与其耐药逆转作用完全相关。更多还原

【Abstract】 This study was aimed to explore the changes of soluble resistance-related calcium binding protein（sorcin）expression in reversion of multidrug resistance of K562/A02 leukemic cell line with defferent concentrations of tetrandrine（Tet）,so as to provide a new theoretic evidence for clinical application of Tet.The inhibition of K562/A02 cell line by daunorubicin（DNR）was assayed by MTT method.The changes of SORCIN gene expression were assayed by RT-PCR.The changes of SORCIN protein expressed were assayed by Western blot.The results showed that Tet could enhance the cytotoxity of DNR to K562/A02 cells（the IC₅0 of DNR+Tet was 11.3±0.17 mg/L,5.15±0.10 mg/L,3.91±0.06 mg/L,and 2.52±0.04 mg/L,when concentrations of Tet were 0 mg/L,0.5 mg/L,1.0 mg/L,and 2.0 mg/L respectively）.The gene encoding sorcin was highly expressed in K562/A02 cells,the expression of which was firstly enhanced in Tet concentration 0.5 mg/L,then attenuated in Tet concentration of 1.0,2.0 mg/L（p<0.05）.Sorcin protein expressed lowly in K562 cells and highly in K562/A02 cells,but the expression of SORCIN protein in K562/A02 cells was enhanced in Tet concentration of 0.5 mg/L,then was attenuated in Tet concentration of 1.0,2.0 mg/L（p<0.05）.It is concluded that the effect of Tet on reversal of K562/A02 drug-resistance shows concentration dependence by MTT assay.Tet reverses multidrug-resistance of K562/A02 cells through regulation of expression of SORCIN gene and protein,but not fully correlates to the reversing effect.更多还原