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目的探讨多药耐药白血病细胞株VLB100提取物-膜微粒介导P-糖蛋白(P-gp)及其编码基因多药耐药基因1(MDR-1)的转移以及对亲代敏感细胞株CCRF-CEM细胞耐药性的影响。方法从VLB100细胞中提取膜微粒,将其与CCRF-CEM细胞共培养。RT-PCR和流式细胞术分别检测MDR-1基因和P-gp的表达,罗丹明123(Rh123)滞留试验检测P-gp功能,MTT法检测膜微粒对阿霉素细胞毒作用的影响。结果膜微粒处理后,CCRF-CEM细胞MDR-1基因及P-gp表达水平增加(P<0.05),细胞内的Rh123积聚减少(P<0.05),细胞对阿霉素的耐药性增加(P<0.05),且均呈剂量依赖性。而多药耐药蛋白1(MRP1)及乳腺癌耐药蛋白(BCRP)基因的表达在处理前后无明显变化(P>0.05)。结论膜微粒可诱导MDR-1基因及P-gp转移,增加CCRF-CEM细胞对阿霉素的耐药性。
Objective To investigate the multidrug resistance leukemia cell line VLB100 extract - membrane-mediated P-glycoprotein (P-gp) and its gene encoding multidrug resistance gene 1 (MDR-1) metastasis and sensitive parental cell line CCRF -CEM cell resistance. Methods Membrane particles were extracted from VLB100 cells and co-cultured with CCRF-CEM cells. The expression of MDR-1 gene and P-gp were detected by RT-PCR and flow cytometry respectively. The function of P-gp was detected by Rh123 retention assay, and the effect of membrane particles on doxorubicin cytotoxicity was detected by MTT assay. Results After treatment with membrane particles, the expression of MDR-1 gene and P-gp in CCRF-CEM cells increased (P <0.05), and the accumulation of Rh123 in cells decreased (P <0.05) and the resistance of cells to doxorubicin increased P <0.05), all in a dose-dependent manner. However, the expression of multidrug resistance protein 1 (MRP1) and breast cancer resistance protein (BCRP) did not change significantly before and after treatment (P> 0.05). Conclusion Membrane particles can induce the MDR-1 gene and P-gp transfer and increase the resistance of doxorubicin to CCRF-CEM cells.