丝氨酸/苏氨酸蛋白激酶抑制剂治疗成人急性白血病的研究

Serine/threonine protein kinase inhibitor in treatment of acute myelogenous leukemia

目的应用基因表达谱芯片来检测丝氨酸/苏氨酸蛋白激酶Raf-1抑制剂对急性髓系白血病(AML)细胞基因表达的影响,试图进一步探讨丝氨酸/苏氨酸蛋白激酶Raf-1抑制剂抗白血病细胞增殖的机制。方法利用cDNA基因芯片研究10例成人AML,用两种不同的荧光素cy3和cy5通过逆转录反应将白血病细胞经丝氨酸/苏氨酸蛋白激酶Raf-1抑制剂作用前后的RNA分别标记成两种探针,并与基因表达谱芯片进行杂交,通过计算机扫描分析得出这些基因在经丝氨酸/苏氨酸蛋白激酶Raf-1抑制剂处理前后的表达差异。结果基因表达谱分析两次重复实验结果,最后筛选出包括与细胞凋亡、细胞周期等相关表达差异的基因共20条,其中12条表达上调,8条表达下调。结论结果提示丝氨酸/苏氨酸蛋白激酶Raf-1抑制剂抗成人急性粒细胞白血病的作用机制与抑制白血病细胞周期和诱导白血病细胞凋亡有很大关系。

Objective The serine/threonine protein kinase （ Raf-1 ） constitutive activation have been identified in a range of human tumors, including acute myelogenous leukemia （AML）. The collective evidence suggests that Raf-1 is a viable anticancer drug target. We therefore targeted Raf-1 downstream of Ras using Raf-1 inhibitor that inhibit Raf-1 kinase activity in the treatment of AML and displayed potent antitumor and antiproliferative activity. The purposes of this study is to investigate the mechanism of anti-leukemia of Raf-1 inhibitor. Methods Experiments were carried out to study 10 adult AML patients by cDNA microarray. Total RNA from AML patients was isolated before and after Raf-1 inhibitor treatment, cDNA probes by RT-PCR were labeled with cy3 and cy5 fluorescence dyes, hybridized with cDNA microarray, and scanned for fluorescent intensity. The resulting images were quantitated by Axon Genepix software. Gene expression profiling after drug treatment were compared to pretreatment. Results Twenty genes related to apoptosis,ce11 cycle and others expressed different after the treatment of Raf-1 inhibitor, 12 genes were up regulated i8 genes down regulated. Conclusions The mechanism of anti-leukemia proliferation of Raf-1 inhibitor is related to cell cycle inhibition and cell apoptosis induced by Raf-1 inhibitor.