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| 细胞自噬在多柔比星诱导淋巴瘤细胞凋亡中的作用 |
| 范佳君1,曾贤2,李玉彬3,王晓丹1,王子玉2,鞠佃文1,2* |
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(1. 同济大学生命科学与技术学院, 上海 200092
2. 复旦大学药学院生物合成教研室,上海 200433
3. 南京农业大学生命科学学院农业部农业环境微生物工程重点开放实验室,南京 210095
*通信作者) |
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| 摘要: |
| 目的 研究细胞自噬在多柔比星诱导Raji细胞发生细胞凋亡过程中所起的作用。方法 以含不同浓度多柔比星的培养液培养Raji细胞,采用蛋白质印迹法观察Raji细胞自噬相关蛋白的表达,并用MDC染色法观察Raji细胞内的自噬体和自噬溶酶体的形成情况；在给予多柔比星的同时使用PI3K通路抑制剂3-MA和自噬溶酶体抑制剂NH4Cl对细胞自噬进行抑制,利用MTT法和流式细胞术测定Raji细胞的细胞活力与凋亡情况。结果 给予多柔比星后,Raji细胞的LC3Ⅱ表达水平和MDC染色荧光强度均高于对照组(P<0.05),证明多柔比星能诱导Raji细胞发生细胞自噬。给予多柔比星后,Raji细胞生长受到抑制,凋亡增加(P<0.01)；用细胞自噬抑制剂3-MA和NH4Cl抑制自噬后,细胞生长受抑和凋亡增加现象更加显著(P<0.05,P<0.01)。结论 在多柔比星杀伤Raji细胞的过程中会诱导细胞发生自噬,抑制自噬能够增强多柔比星对Raji细胞的杀伤作用。 |
| 关键词: 多柔比星 自噬 细胞凋亡 淋巴瘤 |
| DOI：10.3724/SP.J.1008.2012.00595 |
| 投稿时间：2012-03-27修订日期：2012-05-07 |
| 基金项目:科技部新药重大创制项目（2011ZX09102-001-27），上海市重点科技攻关项目(11431920104), 上海市科技人才项目（09XD1421800）. |
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| Roles of autophagy in doxorubicin-induced apoptosis of lymphoma cells |
| FAN Jia-jun1,ZENG Xian2,LI Yu-bin3,WANG Xiao-dan1,WANG Zi-yu2,JU Dian-wen1,2* |
(1. School of Life Science and Technology, Tongji University, Shanghai 200092,China
2. Department of Biosynthesis, School of Pharmacy, Fudan University, Shanghai 200433, China
3. Key Laboratory for Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Science, Nanjing Agricultural University, Nanjing 210095, Jiangsu,China
*Corresponding author.) |
| Abstract: |
| Objective To evaluate the role of autophagy in doxorubicin-induced apoptosis of Raji cells.Methods Raji cells were treated with different concentrations of doxorubicin. Western blotting analysis was used to observe the expression of autophagy-related protein during doxorubicin-induced cell death, and MDC staining was applied to detect autophgosomes and autolysosomes in Raji cells. Cell viability and apoptosis of Raji cells were examined by MTT assay and flow cytometry after the cells were treated with doxorubicin in combination with PI3K pathway inhibitor 3-MA or autolysosome inhibitor NH4Cl. Results Doxorubicin treatment significantly increased LC3Ⅱ expression and the intensity of MDC staining in Raji cells compared with the control group (P<0.05),indicating that doxorubicin can induce autophagy of Raji cells. Compared with the control group, doxorubicin treatment significantly inhibited cell growth and induced apoptosis of Raji cells (P<0.01). PI3K pathway inhibitor 3-MA or autolysosome inhibitor NH4Cl further enhanced the efficacy of doxorubicin in inhibiting cell growth and inducing apoptosis of Raji cells(P<0.05, P<0.01).Conclusion Doxorubicin can induce cell autophagy when killing Raji cells, and inhibition of autophagy can enhance the killing effect of doxorubicin against Raji cells. |
| Key words: doxorubicin autophagy apoptosis lymphoma |
