Li Jing, Chen Bao-An, Zhu Min-Sheng, Gao Fen, Ding Jia-Hua, Gao Chong, Sun Yun-Yu, Cheng Jian, Wang Jun, Zhao Gang, Song Hui-Hui, Ma Yan, Bao Wen, Sun Xin-Cheng, Cheng Hong-Yan, Xu Wen-Lin, Shen Hui-Ling
Department of Hematology, Zhongda Hospital, Southeast University, Nanjing 210009, Jiangsu Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Feb;16(1):65-9.
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.
本研究旨在探讨不同浓度粉防己碱（Tet）对K562/A02白血病细胞株多药耐药逆转过程中可溶性耐药相关钙结合蛋白（sorcin）表达的变化，为粉防己碱的临床应用提供新的理论依据。采用MTT法检测柔红霉素（DNR）对K562/A02细胞株的抑制作用。采用RT-PCR检测SORCIN基因表达的变化。采用Western blot检测SORCIN蛋白表达的变化。结果显示，粉防己碱可增强DNR对K562/A02细胞的细胞毒性（当粉防己碱浓度分别为0 mg/L、0.5 mg/L、1.0 mg/L和2.0 mg/L时，DNR+粉防己碱的IC(50)分别为11.3±0.17 mg/L、5.15±0.10 mg/L、3.91±0.06 mg/L和2.52±0.04 mg/L）。编码sorcin的基因在K562/A02细胞中高表达，其表达在粉防己碱浓度为0.5 mg/L时首先增强，然后在粉防己碱浓度为1.0、2.0 mg/L时减弱（p<0.05）。Sorcin蛋白在K562细胞中低表达，在K562/A02细胞中高表达，但K562/A02细胞中SORCIN蛋白的表达在粉防己碱浓度为0.5 mg/L时增强，然后在粉防己碱浓度为1.0、2.0 mg/L时减弱（p<0.05）。结论：通过MTT法检测，粉防己碱对K562/A02耐药逆转的作用呈浓度依赖性。粉防己碱通过调节SORCIN基因和蛋白的表达逆转K562/A02细胞的多药耐药性，但与逆转效果不完全相关。
