Wang Chun-Ling, Cai Xiao-Hui, Zhang Li-Juan, He Zheng-Mei, Sheng Fei, Cheng Jian, Zhang Yu, Chen Bao-An
Department of Hematology and Oncology, Key Laboratory of Jiangsu Province, Zhongda Hospital, Medical School of Southeast University, Nanjing 210009, Jiangsu Province, China.
Department of Hematology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an 223300, Jiangsu Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Oct;23(5):1318-24. doi: 10.7534/j.issn.1009-2137.2015.05.018.
OBJECTIVE: To explore the effect of magnetic iron nanoparticles ( Fe₃O₄- MNP) in combination with arsenic trioxide and adriamycin on apoptosis and autophagy of Raji cells, a non-Hodgkin's lymphoma (NHL) cell line. METHODS: The growth inhibition rate of Raji cells was analyzed by MTT assay, the cells apoptosis and intracellular concentration of ADM were determined by flow cytometry (FCM), the expression levels of apoptosis-related proteins such as BCL-2, NFκB, Survivin, BAX, P53, and Caspase-3, and related to autophagy-proteins, such as LC3, Beclin-1, and P62/SQSTM1 were detected by Western blot. RESULTS: The growth inhibition of Raji cells in the group of ADM + As₂O₃were higher than that in the group of ADM or As₂O₃alone, however, lower than that in the group of Fe₃O₄- MNP combined with ADM and As₂O₃(ADM+As₂O₃+ MNP) (P < 0.05). The apoptotic rate and accumulation of intracellular ADM in the group of Fe₃O₄- MNP combined with ADM and As₂O₃were significantly higher than those in control, ADM, As₂O₃, and ADM plus As₂O₃groups (P < 0.05). The upregulation of BAX, P53 and Caspase-3 expression and the down regulation of BCL-2, NFκB, and Survivin expression at protein level were more remarkable in the group of ADM+As₂O₃ + MNP, compared with the other groups (P < 0.05). Moreover, the expressions of LC3 and Beclin-1 proteins in the group of ADM+As₂O₃+ MNP were higher, while the expression of P62/SQSTM1 was lower than that in other groups (P < 0.05). CONCLUSION: The Fe3O4 - MNP combined with ADM and As₂O₃can increase the antitumor efficacy on Raji cells by promoting apoptosis and inducing autophagy. It would be a promising strategy for malignant lymphoma therapy.
目的：探讨磁性铁纳米颗粒（Fe₃O₄-MNP）联合三氧化二砷和阿霉素对非霍奇金淋巴瘤（NHL）细胞系Raji细胞凋亡和自噬的影响。 方法：采用MTT法分析Raji细胞的生长抑制率，通过流式细胞术（FCM）检测细胞凋亡及细胞内阿霉素浓度，采用蛋白质免疫印迹法检测凋亡相关蛋白如BCL-2、NFκB、Survivin、BAX、P53和Caspase-３以及自噬相关蛋白如LC3、Beclin-1和P62/SQSTM1的表达水平。 结果：阿霉素＋三氧化二砷组对Raji细胞的生长抑制作用高于阿霉素组或三氧化二砷组，但低于Fe₃O₄-MNP联合阿霉素和三氧化二砷（阿霉素＋三氧化二砷＋MNP）组（P＜0.05）。Fe₃O₄-MNP联合阿霉素和三氧化二砷组的细胞凋亡率及细胞内阿霉素蓄积量均显著高于对照组、阿霉素组、三氧化二砷组及阿霉素加三氧化二砷组（P＜0.05）。与其他组相比，阿霉素＋三氧化二砷＋MNP组在蛋白质水平上BAX、P53和Caspase-３表达上调以及BCL-2、NFκB和Survivin表达下调更为显著（P＜0.05）。此外，阿霉素＋三氧化二砷＋MNP组LC3和Beclin-1蛋白表达较高，而P62/SQSTM1表达低于其他组（P＜0.05）。 结论：Fe₃O₄-MNP联合阿霉素和三氧化二砷可通过促进凋亡和诱导自噬增强对Raji细胞的抗肿瘤疗效。这可能是恶性淋巴瘤治疗的一种有前景的策略。
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