FOXO3a转录因子对白血病HL60细胞增殖和凋亡的影响研究

The research on the effect of transcription factor FOXO3a on proliferation and apoptosis of acute myeloblastic leukemia HL60 cell line

目的:研究转录因子FOXO3a对急性粒细胞白血病HL60细胞株增殖、凋亡的影响及可能的分子机制。方法:将FOXO3a表达质粒pcDNA3.1-FOXO3a及空载对照质粒pcDNA3.1分别经阳离子脂质体介导转染HL60细胞,用G418筛选出FOXO3a稳定表达细胞株。台盼蓝拒染法和MTT法检测HL60细胞增殖,流式细胞仪分析细胞周期和凋亡,RT-PCR及Western blot方法检测基因Bim和bcl-2的表达。结果:筛选到稳定表达FOXO3a基因的细胞株HL60/FOXO3a细胞,与空载体转染组(HL60/empty)及未处理对照组(HL60/untreated)细胞相比,转染组FOXO3a明显抑制HL60细胞生长与增殖,导致G1期阻滞,细胞凋亡明显增加。RT-PCR及Western blot显示FOXO3a明显上调Bim基因的mRNA和蛋白表达,而抑制bcl-2基因mRNA和蛋白的表达。结论:FOXO3a转录因子能抑制HL60细胞的生长和增殖,并诱导其发生凋亡,其机制可能是通过对凋亡相关基因的调控并阻止细胞周期进程来实现的。

Objective：To study the effect of transcription factor FOXO3a on acute myeloblastic leukemia HL60 cell line proliferation,apoptosis and the possible molecular mechanism.Methods： The FOXO3a expression plasmid pcDNA3.1 FOXO3a and the no load control plasmid pcDNA3.1 were separately transfected to HL60 cells mediated by cationic lipid body.The FOXO3a stable expression cell line was screened with G418.HL60 cell proliferation was assayed by Trypan blue exclusion staining and MTT.Cell cycle and apoptosis were analyzed by flow cytometry.The expressions of Bim and bcl-2 genes were detected by RT-PCR and Western blot method.Results： The stable cell lines HL60/FOXO3a cells were screened,which significantly inhibited the growth and proliferation ability of HL60 cells compared with the empty vector transfection group（HL60/empty） and untreated control group（HL60/untreated） cells,and were accompanied with G1 phase block and apoptotic cell increase.The gene FOXO3a significantly up regulated Bim gene mRNA and protein expression,while inhibited the bcl 2 gene mRNA and protein expression.Conclusion： Transcription factor FOXO3a could inhibit HL60 cell growth and proliferation,and induce apoptosis.Its possible mechanism may be involved in the regulation of apoptosis related genes and prevention of cell cycle progression.