Ce^(4+)诱导的花生悬浮细胞程序性死亡

Programmed Cell Death Induced by Ce^(4+) in Suspension Culture of Arachis hypogaea

应用超薄切片透射电镜观察和DNAladder检测技术,对添加不同浓度Ce4+胁迫诱导的花生悬浮细胞进行观测.结果表明,花生悬浮细胞经1 0mmol/LCe4+胁迫诱导培养7d,细胞凋亡率达57 6%;发生凋亡的细胞呈现典型的细胞程序性死亡特征:核仁消失,染色质凝缩、趋边化;线粒体嵴模糊不清,基质电子密度减少,出现大小不一的空泡,有的线粒体缢缩,或破裂,甚至解体;液泡有的破裂,有的相互融合,有的与内质网连接在一起;内质网槽库膨大,并出现自噬泡;DNA特异性降解,产生大小约200bp及其倍数的片断,电泳形成梯状条带.而对照组细胞核DNA完整,呈大片段,细胞器发达,细胞完整.证明Ce4+能诱导花生悬浮细胞发生程序性死亡.

Programmed cell death (PCD) induced by Ce^(4+) with different concentration in suspension culture of Arachis hypogaea was examined, using transmission electron microscopy and DNA laddering. The results suggested that rate of PCD in suspension culture of A. hypogaea induced by 1.0 mmol/L Ce^(4+) at 7 d was 57.6%. And under transmission electron microscopy the following morphological changes were observed: Nucleolus disintegrated, chromatin condensed and margined. The mitochondrion cristea blurred, and mitochondrion full of hollows, decrease of electron density in mitochondrion. The mitochondrion membrane constricted or ruptured, or deteriorated. The vacuoles fused or disrupted, or united with endoplasmic reticulum. The endoplasmic reticulum cisterna dilated, and then formed phagocytic vacuole. In 1.0 mmol/L Ce^(4+)-treated cells at 7 d, agarose gel eletrophoresis revealed that DNA was digested into oligonucleosomal fragments that were times of 200 bp appearing as DNA laddering. Control cells were normal physiological state, they were intact, abundant in organelle and with integral nucleus DNA. The results identified that Ce^(4+) stress could induce PCD in suspension culture of A.hypogaea.