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本研究应用基因芯片探讨酪氨酸激酶抑制剂(STI571)对K562细胞生长、增殖的影响,研究STI571诱导细胞凋亡过程中基因表达的变化及STI571诱导K562细胞凋亡的机制。用RD-PCR技术制备基因芯片探针,然后制成K562细胞表达谱芯片;用相差显微镜观察K562细胞在STI571处理前后的形态变化;用MTT法检测K562细胞的凋亡,用制备的K562细胞表达谱芯片分析基因表达水平。结果表明,在体外培养条件下用1.0μmol/L的STI571处理K562细胞达到一定时间(24小时)后,K562细胞生长明显变缓,出现凋亡细胞的特征。用自制的K562细胞基因表达谱芯片检测显示,STI571诱导K562细胞凋亡前后的基因表达有差异。杂交检测后发现,经STI571诱导后基因表达下调的基因有9个,表达上调的基因有4个。这些表达变化的基因主要包括细胞周期相关基因、细胞代谢通路相关基因、信号转导和转录调节相关基因及抗凋亡基因。结论:STI571能有效抑制K562细胞生长,诱导K562细胞凋亡;筛选获得的靶基因在K562细胞恶性转化过程中发挥了重要作用,这为药物靶基因的筛选提供了理论基础。
In this study, the gene chip was used to investigate the effect of tyrosine kinase inhibitor (STI571) on the growth and proliferation of K562 cells. The gene expression changes during STI571-induced apoptosis and the mechanism of STI571-induced K562 cell apoptosis were investigated. The gene chip probes were prepared by RD-PCR and then were made into K562 cell expression microarray. Morphological changes of K562 cells before and after STI571 treatment were observed by phase-contrast microscopy. The apoptosis of K562 cells was detected by MTT assay and expressed in K562 cells Gene chip analysis of gene expression levels. The results showed that the growth of K562 cells was obviously slowed down and the characteristics of apoptotic cells appeared when K562 cells were treated with 1.0μmol / L STI571 for a certain period of time (24 hours) in vitro. Using self-made K562 cells gene chip microarray detection showed that STI571 induced K562 cells apoptosis before and after the gene expression differences. After hybridization, 9 genes were down-regulated after STI571 induction and 4 genes were up-regulated. These expressed genes mainly include cell cycle related genes, cell metabolic pathway related genes, signal transduction and transcription regulation related genes and anti-apoptotic genes. CONCLUSIONS: STI571 can effectively inhibit the growth of K562 cells and induce the apoptosis of K562 cells. The target gene screened plays an important role in the malignant transformation of K562 cells, which provides a theoretical basis for the screening of target genes.