Rahman Z, Yoshikawa H, Nakajima Y, Tasaka K
Department of Parasitology and Immunology, Yamanashi Medical University, 1110 Shimokato, Tamaho-cho, 409-3898, Yamanashi, Japan.
Immunol Lett. 2001 Jan 15;75(3):199-208. doi: 10.1016/s0165-2478(00)00322-9.
Here we report, interleukin-6 (IL-6) dependent mouse B-cell hybridoma, 7TD1 cells underwent apoptotic cell death with the starvation of IL-6. First, 7TD1 cells cultured without IL-6 arrested at G0/G1 phase (maximum accumulation at 24 h ) of the cell cycle. After that, the parameters of apoptosis namely, decreased mitochondrial transmembrane potential (DeltaPsi(m)), activation of caspases, DNA fragmentation and morphological changes (condensed nucleus and formation of apoptotic bodies) were observed. As evidents by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses, down-regulation of Pim-1 (a serine/threonine kinase) and Bcl-2 was observed in the IL-6-depleted 7TD1 cells. There was no change in the expression of c-Myc, Bcl-xL and Mcl-1, even at 48 h of IL-6-depletion. Taken together, these results indicate that IL-6 withdrawn from the 7TD1 cells resulted in G0/G1 arrest and then caspase-dependent apoptosis via mitochondrial pathway by down-regulation of Pim-1 and Bcl-2, which may be essential for anti-apoptotic signals of IL-6.
在此我们报告,依赖白细胞介素-6(IL-6)的小鼠B细胞杂交瘤7TD1细胞在IL-6缺乏时会发生凋亡性细胞死亡。首先,在无IL-6的情况下培养的7TD1细胞停滞在细胞周期的G0/G1期(24小时时积累达到最大值)。之后,观察到凋亡的参数,即线粒体跨膜电位降低(ΔΨm)、半胱天冬酶激活、DNA片段化以及形态学变化(细胞核浓缩和凋亡小体形成)。通过逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹分析表明,在缺乏IL-6的7TD1细胞中观察到Pim-1(一种丝氨酸/苏氨酸激酶)和Bcl-2的下调。即使在缺乏IL-6 48小时时,c-Myc、Bcl-xL和Mcl-1的表达也没有变化。综上所述,这些结果表明,从7TD1细胞中去除IL-6会导致G0/G1期停滞,然后通过下调Pim-1和Bcl-2,经由线粒体途径发生半胱天冬酶依赖性凋亡,这可能是IL-6抗凋亡信号所必需的。