摘要
目的:观察甲磺酸伊马替尼干预K562细胞后SHIP、caspase-1、caspase-3、caspase-9及bcl-2基因表达的变化,初步探讨伊马替尼促凋亡的作用途径。方法:将不同浓度甲磺酸伊马替尼与K562细胞在体外共培养,于不同时段留取标本,采用实时定量PCR方法检测SHIP基因表达水平的变化,并用半定量逆转录PCR方法检测抗凋亡基因bcl-2及促凋亡基因caspase-1、caspase-3、caspase-9表达水平的变化,用MTT法观察伊替尼对细胞增殖的抑制作用,用膜联蛋白Ⅴ/碘化丙锭(AnnexinⅤ/PI)双标记法分析细胞凋亡。结果:甲磺酸伊马替尼可使K562细胞内SHIP基因表达水平升高,且与剂量和作用时间有关;caspase-9表达水平亦升高并与SHIP基因表达水平有直线相关关系;而caspase-1、caspase-3和bcl-2表达水平无显著变化;伊马替尼可使细胞增殖率降低、凋亡率增加,并可见到明显的形态学变化。结论:甲磺酸伊马替尼可使K562细胞内SHIP及caspase-9基因表达水平升高且可促进细胞凋亡,其促凋亡作用机制可能与上调SHIP和caspase-9基因表达有关。
Objective： To observe the changes of SHIP,caspase-1, caspase-3, caspase-9 and bcl-2 gene expression levels in K562 cells after mesylate imatinib （MI） treatment and explore the possible mechanism for apoptosis-inducing effects of imatinib, Methods： K562 cells were cultured with MI at different concentrations. The cells were collected at different time points. Real-time quantitative PCR was used to detect the expression level of SHIP gene. Semi-quantitative reverse transcriptase PCR was used to detect the mRNA transcription of anti-apoptotic gene bcl-2 and pro-apoptotic genes caspase-1, caspase-3 and caspase-9. The cell proliferation was measured by MTT assay. The cell apoptosis was analyzed by Annexin V/PI double staining flow cytometry. Results： MI significantly increased the expression of SHIP gene in a time-and dose-dependent manner. Caspase-9 gene was also up-regulated after MI treatment and had linear correlation with SHIP gene expression. The expression levels of caspase-1, caspase-3 and bcl-2 had no significant changes. MI down-regulated the proliferation and induced the apoptosis of K562 cells. The morphological changes of typical of apoptosis were observed. Conclusion： MI significantly increases the expressions of SHIP gene and caspase-9 gene and induces apoptosis of K562 cells. The mechanism for the apoptosis-inducing effects of imatinib may be associated with the up-regulation of SHIP and caspase-9 gene.
出处
《肿瘤》
CAS
CSCD
北大核心
2007年第11期853-856,共4页
Tumor
基金
国家自然科学基金资助项目(编号:30240011)
