关迪,崔韶晖,杨珺,张传敏,支德福,张树彪
GUAN Di,CUI Shao-hui,YANG Jun,ZHANG Chuan-min,ZHI De-fu,ZHANG Shu-biao

• 1:大连工业大学生物工程学院
• 2:大连民族学院生命科学学院

摘要(Abstract)：

目的研究DDCTMA阳离子脂质体对体外培养的人喉癌细胞系(Hep-2)细胞增殖和凋亡作用影响,进而分析其细胞毒性作用机制。方法采用MTT法检测不同质量浓度DDCTMA阳离子脂质体作用下的细胞存活率,据此选择适当质量浓度的DDCTMA阳离子脂质体分别处理Hep-2细胞,使用活细胞/凋亡细胞/坏死细胞鉴别试剂盒、Hoechst 33258检测法及TUNEL原位测定法进行凋亡细胞形态学鉴定。定量检测酶caspase-3和caspase-9的酶活力变化。结果随着DDCTMA阳离子脂质体质量浓度的增大,细胞存活率逐渐下降;活细胞减少,坏死和凋亡细胞数量都增加;对细胞核的染色能观察到凋亡细胞数量呈倍数增加,呈现浓度依赖性;caspase-3和caspase-9的酶活力也有相应增高。结论 DDCTMA阳离子脂质体对Hep-2细胞的毒性作用表现是:DDCTMA阳离子脂质体质量浓度在9~48mg·L-1内主要诱导部分细胞凋亡,质量浓度超过48mg·L-1主要引起细胞坏死。
Objective To study the cytotoxicity mechanism of DDCTM A cationic liposome against human throat epidermis cancer cells(Hep-2) in vitro.Methods The M TT method was used to detect cell survival rate with different concentrations of DDCTM A cationic liposome.After the proper concentrations of DDCTM A cationic liposome were selected to treat Hep-2 cells,live cell / apoptosis / necrosis cell identification kits,Hoechst 33258 assay and TUNEL apoptotic cells identification kits were applied to stain the cells,which were observed by inverted fluorescence microscope.Quantitative detection of the enzymatic activity of caspase-3 and caspase-9 was performed.Results With the increase of DDCTM A cationic liposome concentration,the survival rate of Hep-2 cells gradually decreased,and necrosis and apoptosis cells increased.The enzymatic activity of caspase-3 and caspase-9 increased accordingly.Conclusions DDCTM A cationic liposome with concentrations of 9 μg·m L- 1- 48 mg·L- 1mainly induced apoptosis,more than 48 mg·L- 1mainly caused cell death of Hep-2 cells.

关键词(KeyWords)： 阳离子脂质体;基因转运;细胞毒性;细胞坏死;细胞凋亡
cationic liposome;gene delivery;cytotoxicity;necrosis;apoptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(21176046);; 中央高校基本科研业务费专项资金项目(DC12010142,DC12010104)

作者(Author): 关迪,崔韶晖,杨珺,张传敏,支德福,张树彪
GUAN Di,CUI Shao-hui,YANG Jun,ZHANG Chuan-min,ZHI De-fu,ZHANG Shu-biao

DOI: 10.14066/j.cnki.cn21-1349/r.2015.02.009

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