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本研究旨在探讨汉防己甲素(TET)对人慢性髓系白血病(CML)急性红白血病细胞株K562/A02多药耐药的逆转作用,及其逆转耐药与细胞周期依赖性激酶抑制因子(p21)、P糖蛋白(P-gp)及其基因的关系,为TET的临床应用提供新的理论基础。K562/A02细胞实验分组为:①单用柔红霉素(DNR)对照组;②DNR+TET0.5mg/L组;③DNR+TET1.0mg/L组;④DNR+TET2.0mg/L组。采用Westernblot法检测实验各组K562/A02细胞P21的表达。采用流式细胞仪(FCM)检测实验各组细胞P-gp的表达,采用半定量PCR(RT-PCR)测定细胞MDR1基因mRNA表达水平。结果表明:K562/A02细胞中P21蛋白的表达随着TET浓度的增加而增强,P-gp蛋白的表达随着TET浓度的增加而减低。K562细胞中mdr1基因几乎无表达,K562/A02细胞中mdr1基因高表达,随着TET浓度的增加K562/A02细胞mdr1基因表达不断降低。结论:TET对K562/A02耐药具有逆转作用且呈浓度依赖性,其逆转耐药可能与其上调P21蛋白的表达、下调mdr1及P-gp的表达有关。
This study was designed to investigate the reversal effect of tetrandrine (TET) on multidrug resistance in acute myelogenous leukemia cell line K562 / A02 in human chronic myelogenous leukemia (CML) and to investigate its reversal of drug resistance and cell cycle dependent kinase inhibitor (p21), P-glycoprotein (P-gp) and their genes, providing a new theoretical basis for the clinical application of TET. K562 / A02 cells were divided into groups as follows: ① single daunorubicin (DNR) control group; ② DNR + TET0.5mg / L group; ③DNR + TET1.0mg / L group; ④DNR + TET2.0mg / L group. Western blot was used to detect the expression of P21 in K562 / A02 cells. The expression of P-gp in each group was detected by flow cytometry (FCM), and the mRNA expression of MDR1 was detected by semi-quantitative PCR (RT-PCR). The results showed that the expression of P21 protein in K562 / A02 cells increased with the increase of TET concentration, and the expression of P-gp protein decreased with the increase of TET concentration. There was almost no expression of mdr1 gene in K562 cells and high expression of mdr1 gene in K562 / A02 cells. The expression of mdr1 gene in K562 / A02 cells was decreased with the increase of TET concentration. CONCLUSION: TET has a reversal effect on K562 / A02 resistance in a concentration-dependent manner. The reversal of drug resistance may be related to its up-regulation of P21 protein expression and down-regulation of mdr1 and P-gp expression.