汉防己甲素对K562/A02细胞株SORCIN基因表达的影响

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。

Influence of Tetrandrine on SORCIN Gene Expression in K562/A02 Cell Line

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 different 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 cytotoxicity of DNR to K562/A02 cells (the IC(50) 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.