Luo D, Cheng S C, Xie Y
Department of Biology, Hong Kong University of Science and Technology, SAR, China.
Br J Biomed Sci. 1999;56(2):114-22.
This study demonstrates that two anticancer drugs, taxol and doxorubicin (Dox), can kill human hepatoblastoma HepG2 cells in a dose-dependent manner via the induction of apoptosis. Characteristic events, including externalization of phosphatidylserine, cytoplasmic shrinkage, chromatin condensation and DNA degradation, were observed in a large majority of the drug-treated cells. DNA fragmentation showed that a ladder of DNA fragments of approximately 200 bp multiples was observed in taxol-treated, but not in Dox-treated, cells. In addition, the expression patterns of Bcl-2 family members during taxol or Dox treatment were investigated. Results from Western blot analysis indicated that HepG2 cells did not express either the death repressor Bcl-2, or the death promoters Bcl-XS and Bax. However, during the apoptotic process one death repressor, Bcl-XL, and two death promoters, Bak and Bad, were expressed. The expression levels of Bcl-XL and Bak remained unchanged, whereas the level of Bad was down-regulated. As the ratio between death repressors and death promoters in the Bcl-2 family will determine the sensitivity of cells to apoptotic stimuli, the findings suggest that the changed expression patterns of Bcl-2 family proteins caused by anticancer drugs in liver cancer cells may be involved in chemoresistance.
本研究表明,两种抗癌药物紫杉醇和阿霉素(Dox)可通过诱导凋亡以剂量依赖方式杀死人肝癌细胞HepG2。在大多数经药物处理的细胞中观察到了包括磷脂酰丝氨酸外化、细胞质收缩、染色质凝聚和DNA降解在内的特征性事件。DNA片段化显示,在紫杉醇处理的细胞中观察到约200 bp倍数的DNA片段梯状条带,而在阿霉素处理的细胞中未观察到。此外,还研究了紫杉醇或阿霉素处理期间Bcl-2家族成员的表达模式。蛋白质印迹分析结果表明,HepG2细胞既不表达死亡抑制因子Bcl-2,也不表达死亡促进因子Bcl-XS和Bax。然而,在凋亡过程中,一种死亡抑制因子Bcl-XL以及两种死亡促进因子Bak和Bad表达。Bcl-XL和Bak的表达水平保持不变,而Bad的水平下调。由于Bcl-2家族中死亡抑制因子与死亡促进因子之间的比例将决定细胞对凋亡刺激的敏感性,这些发现表明,抗癌药物在肝癌细胞中引起的Bcl-2家族蛋白表达模式变化可能与化疗耐药性有关。