放线菌素D诱导的SL-1细胞凋亡的初步研究

Preliminary Studies on Apoptosis Induced by Actinomycin D in Spodoptera Litura Cells

研究了一定浓度的放线菌素D诱导SL-1细胞凋亡作用。以终浓度为50 ng/mL,100ng/mL,200 ng/mL,500 ng/mL的放线菌素D处理正常SL-1细胞,24 h后,终浓度为200 ng/mL,500 ng/mL的放线菌素D处理的细胞出现较明显的细胞凋亡现象,光学显微镜下可见细胞膜表面突出或形成小泡,细胞碎裂成凋亡小体;DAPI染色荧光显示细胞核逐渐形变,直至碎裂成小块而被凋亡小体包裹;DNA琼脂凝胶电泳呈典型的凋亡细胞DNA梯形条带;流式细胞仪检测结果,对照和以上四种浓度的放线菌素D处理的细胞凋亡率分别为3.43%,10.61%,15.60%,32.67%,72.24%。实验结果证明一定浓度的放线菌素D能诱导SL-1细胞凋亡,且成剂量正相关。

That actinomycin D at certain concentrations could induce SL-1 cells apoptosis was studied. SL-1 cells were treated with final concentrations of 50ng/mL, 100ng/mL, 200ng/mL,500ng/mL actinomycin D, and it was clearly observed 24 hours after treatment, the addition of actinomycin D at final concentrations of 200ng/mL, 500ng/ml to induce apoptosis, microscopic examination of SL-1 cell treated with ActD reaveal progressive blebbing and culminating in most cell destruction. The fragmentation of the cell nuclei is shown with DAPI. Agarose gel eleetrophoresis of the DNA extrate from cells treated with ActD shows typical DNA ladder. Control and the cells are treated with 50ng/ml, 100ng/mL, 200ng/mL,5OOng/mL actinomycin D for 24 h the percentage of apoptotic cells increased from 3.34%, 10.36%, 15.66%, 31.68%, to 74.75% , which is analyzed by flow cytometry. This suggests actinomycin D at certain concentrations could induce SL-1 cells apoptosis, which is much dependent on the concentration of dose of actinomycin D.