苏云金芽孢杆菌Bt9875晶体蛋白诱导HL-60细胞凋亡

目的]研究苏云金芽孢杆菌(Bacillus thuringiensis,Bt)Bt9875菌株晶体蛋白对人急性髓细胞性白血病细胞HL-60的影响.方法]采用MTT比色、荧光显微观察、DNA凝胶电泳、流式细胞术等方法来检测不同浓度的Bt9875晶体蛋白处理后HL-60细胞的凋亡特征.结果]Bt9875晶体蛋白对HL-60细胞的生长具有明显的抑制作用,且随着蛋白质浓度的增加对HL-60细胞生长抑制愈加明显,而对正常人外周血单个核细胞(PBMC)无作用;荧光显微镜下观察发现经该蛋白作用后HL-60细胞核的形态呈现凋亡特征;流式细胞术分析表明,HL-60细胞经100 μg/mL晶体蛋白作用后,凋亡率达到52%;琼脂糖凝胶电泳显示细胞DNA呈梯状降解.结论]初步证明了Bt9875晶体蛋白在体外能够明显抑制HL-60细胞的增长,并诱导其凋亡,这为苏云金芽抱杆菌晶体蛋白的应用开创了新的思路.

Apoptosis of HL-60 cells induced by crystal proteins from Bacillus thuringiensis Bt9875

Objective] To study the effect of Bt9875 crystal protein treated with proteinase K on human cancer cells, HL-60. Methods] We used the methods of Thiazolyl Blue Tetrazolium Bromide, fluorescence microscopy examination, agarose gel electrophores and flow cytometry to detect the growth inhibition rate and apoptosis characters of the HL-60 cells that were treated with different concentration of Bt9875 crystal protein. Results] Bt9875 crystal protein inhibited the growth of HL-60 cells evidently in a dose-dependent manner, with minimal effects on normal human peripheral blood mononuclear cells (PBMCs). The nuclei of HL-60 cells showed the characteristics of apoptosis. The analysis by flow cytometry indicated that the apoptosis rate of HL-60 cells was 52% after treatment with Bt9875 crystal protein (100 mg/mL). DNA analyzed by agarose gel electrophoresis showed "ladder" pattern. Conclusion] Bt9875 crystal protein could inhibit the growth of HL-60 and induced its apoptosis, which provided a foundation for use of Bt9875 crystal protein to cure the acute myeloid leukemia.