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目的阐明PHⅡ-7逆转肿瘤耐药的机制。方法用MTT法测定阿霉素、PHⅡ-7及二者联合用药对人白血病细胞系K562及其多药耐药细胞系K562/A02的IC50值,提取不同浓度PHⅡ-7作用48h后K562和K562/A02细胞RNA,以实时定量PCR的方法分析PHⅡ-7对MDR1基因表达水平的影响,最后通过激光共聚焦显微镜和流式细胞仪观察PHⅡ-7作用前后,K562和K562/A02细胞内阿霉素浓度的改变。结果 PHⅡ-7本身具有抗肿瘤活性,对K562和K562/A02IC50值分别为(1.37±0.37)μmol·L-1;(1.48±0.34)μmol·L-1。此外PHⅡ-7还能协同提高阿霉素的细胞毒作用,K562组CDI值为0.22,K562/A02组CDI值为0.09,其对耐药细胞的协同作用更为明显。阿霉素和PHⅡ-7同时作用于K562/A02,随着PHⅡ-7作用剂量的增加,K562/A02细胞MDR1基因表达水平逐渐下降;细胞内阿霉素浓度伴随PHⅡ-7作用时间的延长而逐渐提高。结论 PHⅡ-7不仅本身是有效的肿瘤细胞化疗药物,还具有下调MDR1耐药基因表达的作用,从而有效逆转K562/A02细胞的耐药现象,增强化疗药物阿霉素的细胞杀伤作用。
Objective To clarify the mechanism of PHⅡ-7 reversing tumor drug resistance. Methods IC50 values of doxorubicin, PHⅡ-7 and their combination therapy on human leukemia cell line K562 and multidrug-resistant cell line K562 / A02 were determined by MTT assay. K562 and K562 cells were treated with different concentrations of PHⅡ-7 for 48h / A02 cells by real-time quantitative PCR method to analyze the effect of PHⅡ-7 on MDR1 gene expression. Finally, the expression of MDR1 gene in K562 and K562 / A02 cells was observed by laser scanning confocal microscope and flow cytometry. Change in concentration. Results PHⅡ-7 itself had anti-tumor activity, and the values of K562 and K562 / A02IC50 were (1.37 ± 0.37) μmol·L-1 and (1.48 ± 0.34) μmol·L-1, respectively. In addition, PHII-7 can synergistically enhance the cytotoxicity of doxorubicin. The CDI value of K562 group was 0.22, and the CDI value of K562 / A02 group was 0.09. The synergistic effect of PHII-7 on drug-resistant cells was more obvious. Adriamycin and PHⅡ-7 act on K562 / A02 at the same time. With the increasing dose of PHⅡ-7, the expression of MDR1 gene in K562 / A02 cells gradually decreased. With the prolongation of PHⅡ-7 Gradually increase. Conclusion PHⅡ-7 is not only an effective chemotherapeutic agent for tumor cells, but also has the effect of down-regulating the expression of MDR1 resistance gene, thereby effectively reversing the drug resistance of K562 / A02 cells and enhancing the cell killing effect of chemotherapeutic drug adriamycin.