Genistein induces apoptosis in human hepatocellular carcinomas via interaction of endoplasmic reticulum stress and mitochondrial insult

Biochem Pharmacol. 2007 Mar 15;73(6):782-92. doi: 10.1016/j.bcp.2006.11.027. Epub 2006 Dec 3.


Hepatocellular carcinoma is a very common malignancy and is chemoresistant to currently available chemotherapeutic agents. Endoplasmic reticulum (ER) stress-induced apoptotic pathway is suggested to be less affected by the resistance mechanisms, becoming a potential target of chemotherapeutic strategy. The anticancer effects and expression of GADD153, a transcription factor induced by ER stress, were examined in hepatocellular carcinoma Hep3B cells. The correlation between these two parameters was constructed under flavonoid stimulation with a correlation coefficient (r) of 0.8. The data also showed that genistein (isoflavone) was the most effective one. Genistein induced the activation of several ER stress-relevant regulators, including m-calpain, GADD153, GRP78 and caspase-12. Furthermore, genistein-induced effect was inhibited in cells transfected with antisense GADD153 cDNA, indicating a functional role of GADD153. Notably, genistein induced the activation of caspase-2, whereas did not cause the DNA damage. It also triggered the production of ROS. The antioxidant trolox significantly reduced ROS accumulation, but did not modify genistein-induced apoptotic cell death. The long-term exposure (48 h) of cells to genistein caused Mcl-1 down-regulation and Bad cleavage; furthermore, cyclosporin A (an inhibitor of mitochondrial permeability transition pore) almost completely abolished genistein-induced loss of mitochondrial membrane potential, and induced a 30% reverse of apoptosis caused by long-term treatment (48 h) of genistein, suggesting the involvement of mitochondrial stress in the late phase of genistein-induced effect. Taken together, it is suggested that genistein induces the anticancer effect through a mechanism initiated by ER stress and facilitated by mitochondrial insult in Hep3B cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects*
  • Apoptotic Protease-Activating Factor 1 / physiology
  • Carcinoma, Hepatocellular / drug therapy*
  • Carcinoma, Hepatocellular / pathology
  • Caspases / physiology
  • Cell Line, Tumor
  • Endoplasmic Reticulum / drug effects*
  • Endoplasmic Reticulum / metabolism
  • Genistein / pharmacology*
  • Heat-Shock Proteins / genetics
  • Heat-Shock Proteins / physiology
  • Humans
  • Liver Neoplasms / drug therapy*
  • Liver Neoplasms / pathology
  • Mitochondria / drug effects*
  • Mitochondria / metabolism
  • Molecular Chaperones / genetics
  • Molecular Chaperones / physiology
  • Reactive Oxygen Species / metabolism
  • Transcription Factor CHOP / genetics
  • Transcription Factor CHOP / physiology


  • APAF1 protein, human
  • Antineoplastic Agents
  • Apoptotic Protease-Activating Factor 1
  • DDIT3 protein, human
  • Heat-Shock Proteins
  • Molecular Chaperones
  • Reactive Oxygen Species
  • Transcription Factor CHOP
  • Genistein
  • Caspases
  • molecular chaperone GRP78