摘要
目的:明确低氧预适应诱发胶质瘤细胞系SK-N-SH对BCNU的耐药,研究在低氧预适应的过程中一氧化氮合酶的作用及其机制。方法:用300μmol/L CoCl2预处理SK-N-SH细胞3小时,24小时后以BCNU刺激细胞,MTT法测定细胞增殖,流式细胞技术测定细胞的凋亡。RT-PCR技术检测神经型一氧化氮合酶和凋亡相关基因的表达情况。结果:300μmol/L CoCl2预处理可以明显增加细胞在BCNU条件下的增殖能力,减少凋亡。预适应可以上调神经型一氧化氮合酶的表达,同时可以上调凋亡抑制基因Bcl-2的表达,下调凋亡促进基因Apaf-1的表达。这些调节作用可被神经型一氧化氮合酶的抑制剂7-NI抑制。结论:300μmol/L CoCl2预处理SK-N-SH细胞3小时可以增加肿瘤细胞对BCNU的耐药作用。神经型一氧化氮合酶在预适应中可调节Bcl-2和Apaf-1基因的表达从而起到重要的作用。
Objective： To confirm that BCNU resistance is induced by hypoxic preconditioning and to clarify the role of neuronal nitric oxide synthase （nNOS） in hypoxic preconditioning. Methods： SK-N-SH cells were pretreated using 300 μM CoCl2 for 3 h, then 24 hours later the cells were treated with 30？g/ml BCNU. MTF assay was used to detect cell proliferation and FACS was used to determine the apoptotic index. RT-PCR was used to detect expression of nNOS and other apoptosis-related genes after hypoxic preconditioning. Results： Pretreatment with 300 μM COCl2 for 3 h followed by treatment with BCNU 24 hours later significantly increased proliferation and decreased the apoptotic index compared to BCNU treatment without hypoxic pretreatment. Expression of nNOS was upregulated after preconditioning, while expression of the Bcl-2 and Apaf-1 genes was upregulated and downregulated, respectively. These changes can be arrested by 7-NI, an inhibitor of nNOS. Conclusion： The 3-h hypoxic pretreatment can induce BCNU resistance in glioma cells in vitro. During the preconditioning, nNOS can regulate the expression of the Bcl-2 and Apaf-1 gene, thus playing an important role.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2007年第6期301-304,共4页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金资助(编号:30400463)
关键词
低氧预适应
神经型一氧化氮含酶
凋亡
化疗耐药
Hypoxic preconditioning
Neuronal nitric oxide synthase
Apoptosis chemoresistance
