摘要
目的:探讨细胞色素C在体外作用于HL-60细胞时细胞发生的变化及其相关凋亡基因bcl-2、bcl-xl表达变化的机制。方法:用不同浓度的细胞色素C作用于HL-60细胞24h,然后用MTT检测细胞色素C对HL-60细胞抑制率;用普通光镜、荧光显微镜检测HL-60细胞形态的变化;用流式细胞仪、DNA凝胶电泳对HL-60细胞凋亡的检测;用RT-PCR检测bcl-2、bcl-xlmRNA表达的变化。结果:细胞抑制率随着细胞色素C浓度的增加而增加;当细胞色素C浓度在0-37·5mg/L作用HL-60细胞24h,随着细胞色素C浓度的增加,HL-60细胞发生的凋亡逐渐增加,可见典型的凋亡细胞和明显的DNA梯度条带;同时,在该浓度范围内,bcl-2、bcl-xlmRNA表达逐渐减少;当细胞色素C浓度大于37·5mg/L时,细胞凋亡率并不增加,而是下降,但是坏死细胞明显增加。结论:一定浓度细胞色素C能诱导HL-60细胞发生凋亡,并且细胞凋亡率、bcl-2、bcl-xl基因表达的变化与细胞色素C浓度呈一定的量效依赖关系,细胞色素C诱导HL-60细胞凋亡可能与抑制bcl-2、bcl-xl基因的表达有一定的关系。
AIM ： To study the effect of cytochrome C on HL - 60 cells in vitro and the expression of relevant apoptotic genes. METHODS ： HL - 60 cells were treated with different concentrations of cytochrome C for 24 h. The suppressing rate was assayed by MTT. The morphology of cell was observed by microscope and fluorescence microscope. The apoptosis was assayed by flow cytometry （FCM） and DNA electrophoresis. The expression changes of bcl - 2 and bcl - xl mRNA was examined by RT- PCR. RESULTS： The suppressing rate increased with the increase in the cytochrome C concentrations. When treated with 0 - 37. 5 mg/L cytochrome C for 24 h, the percentage of apoptotic HL - 60 cells increased in a dose - dependent manner, and the typical cells and the appearance of apoptotic DNA ladder were observed. At the same time, within this range of concentration, the expression of bcl - 2 and bcl - xl mRNA decreased gradually. When treated with cytochrome C at concentration higher than 37.5 mg/L, the percentage of apoptotic HL - 60 cells did not increase, but decreased, while the cell necrosis was observed. CONCLUSIONS ： It suggested from the results that at certain range of conentration, cytochrome C induces apoptosis or necrosis in HL - 60 cells. The percentage of apoptosis, the changes of expression of bcl- 2 and bcl -xl depend on the dose of cytochrome C. The mechanism that cytochrome C induces apoptosis in HL - 60 cells may be related to suppressing the expression of bcl - 2 and bcl - xl.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第9期1729-1733,共5页
Chinese Journal of Pathophysiology
基金
广东省卫生厅课题资助(No.2002501)
