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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。
This study was designed to investigate the relationship between tetrandrine (Tet) and K562 / A02 cell resistance and SORCIN gene expression, and provide a new theoretical basis for the clinical application of Tet. Tetracycline (Tet) was selected by MTT method. After incubated with K562 / A02 cell line for 48 hours, the effect of Tet on cytotoxicity of daunorubicin (DNR) was detected by MTT assay. The expression of SORCIN gene was detected by RT-PCR The protein expression of sorcin was detected by Western blot. The results showed that Tet could enhance the cytotoxicity of DNR to K562 / A02 at the concentration range selected (IC50 of DNR and Tet at Tet concentrations of 0, 0.5, 1.0 and 2.0 mg / L were 11.3 ± 0.17 and 5.15 ± 0.10,3.91 ± 0.06,2.52 ± 0.04mg / L, p <0.01). The expression of SORCIN gene in K562 / A02 cells was high, and the expression of SORCIN first increased and then decreased with the increase of Tet concentration (Tet concentration was 0.5mg / L, enhanced at 1.0 and 2.0 mg / L, p <0.05); sorcin protein was low in K652 cells and sorcin protein was highly expressed in K562 / A02 cells, and then increased and then decreased with the increase of Tet concentration Increased at 0.5 mg / L, weakened at 1.0 and 2.0 mg / L, p <0.05). CONCLUSION: MTT showed that Tet could reverse the drug resistance of K562 / A02 in a concentration-dependent manner. Tet may be involved in reversing the drug resistance of K562 / A02 by adjusting the expression of SORCIN gene and protein, but not with its drug resistance reversal effect.