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目的研究Mda-7/IL-24对B细胞淋巴瘤化疗敏感性的影响并探讨其作用机制。方法构建稳定过表达Mda-7/IL-24的人B表型淋巴瘤Raji和Daudi细胞株,MTS法检测稳定转染Mda-7/IL-24对Raji和Daudi细胞在顺铂、表柔比星及长春新碱作用下化疗敏感性的影响,并计算IC_(50)值;流式细胞术(flow cytometry,FCM)检测细胞凋亡水平及胞内罗丹明-123(rhodamine-123)聚集量;RT-PCR及Western blot技术分别从mRNA及蛋白水平检测转染Mda-7/IL-24对,Raji和Daudi细胞表达耐药相关基因MDR1、MRP1及BMI1水平的影响。Western blot技术检测转染Mda-7/IL-24对Raji和Daudi细胞总ERK1/2及p-ERK1/2表达水平的影响。结果在CDDP、epirubicin及VCR作用下,过表达Mda-7/IL-24的Raji和Daudi细胞株增殖水平明显下降,化疗敏感性明显增高,而化疗药物IC_(50)值明显下降,同时,细胞凋亡水平明显增加;过表达Mda-7/IL-24的Raji和Daudi细胞肿瘤耐药相关基因MDR1、MRP1、BMI1、总ERK1/2及p-ERK1/2的表达水平均明显下降,而胞内Rhodamine-123聚集量明显增加。结论转染Mda-7/IL-24可能通过调节ERK途径的活化改变肿瘤多药耐药相关基因的表达,从而促进B细胞淋巴瘤对化疗药物的敏感性。因此Mda-7/IL-24在B细胞淋巴瘤的治疗中可能具有防治多药耐药的作用。
Objective To investigate the effect of Mda-7 / IL-24 on chemosensitivity of B-cell lymphoma and its mechanism. Methods Raji and Daudi cell lines stably expressing Mda-7 / IL-24 were constructed. MTS-positive cells were stained with cisplatin and epirubicin (50) values were determined by flow cytometry (FCM), and the concentration of rhodamine-123 in the cells was detected by flow cytometry (FCM) The expression of MDR1, MRP1 and BMI1 in Raji and Daudi cells were detected by RT-PCR and Western blot respectively from the mRNA and protein levels of Mda-7 / IL-24. Western blot was used to detect the expression of ERK1 / 2 and p-ERK1 / 2 in Raji and Daudi cells transfected with Mda-7 / IL-24. Results Under CDDP, epirubicin and VCR, the proliferation of Raji and Daudi cells over-expressing Mda-7 / IL-24 was significantly decreased, the chemosensitivity was significantly increased, while the IC 50 value of chemotherapeutic drugs was significantly decreased. The level of apoptosis was significantly increased; the expression of MDR1, MRP1, BMI1, total ERK1 / 2 and p-ERK1 / 2 in Raji and Daudi cells overexpressing Mda-7 / IL- Rhodamine-123 aggregates increased significantly. Conclusion Transfection of Mda-7 / IL-24 may change the expression of multidrug resistance-related genes by regulating the activation of ERK pathway, thereby promoting the sensitivity of B-cell lymphoma to chemotherapeutic drugs. Therefore, Mda-7 / IL-24 may play a role in the prevention and treatment of multidrug resistance in the treatment of B cell lymphoma.