title: Hypocrellin B in hepatocellular carcinoma cells: Subcellular localization and sonodynamic damage.
abstract: OBJECTIVE
  To study subcellular localization of hypocrellin B in hepatocellular carcinoma cells, and hypocrellin B-mediated sonodynamic action-induced cell damage.
  
  
  METHODS
  After incubation with 2.5 μM of hypocrellin B, human hepatocellular carcinoma HepG2 cells were exposed to ultrasound waves for 8 sec at an intensity of 0.46 W/cm(2). Clonogenic survival of HepG2 cells was measured using a colony forming assay and light microscope. Ultrastructural morphology was observed using transmission electron microscope (TEM) and mitochondrial membrane potential (MMP) was assessed using confocal laser scanning microcope (CLSM) after rhodamine 123 staining. Additionally, subcellular localization of hypocrellin B in HepG2 cells with organelle probe staining was also observed using CLSM.
  
  
  RESULTS
  The colony forming units of HepG2 cells decreased substantially after sonodynamic treatment. The results of TEM showed microvilli disappearance, apoptotic body formation, swollen mitochondria with loss of cristae and mitochondrial myelin-like features (or membrane whorls). Collapse of MMP was found in the treated cells. Hypocrellin B was distributed in mitochondria and lysosomes as well as in endoplasmic reticulum and Golgi apparatus.
  
  
  CONCLUSIONS
  The findings demonstrated that sonodynamic action of hypocrellin B induced mitochondrial damage, survival inhibition, and apoptosis of HepG2 cells. Additionally, other subcellular organelles such as endoplasmic reticulum, Golgi apparatus and lysosomes were also the targets of hypocrellin B-mediated sonodynamic action as well as mitochondria.
authors.affiliation: Department of Ultrasound Medicine, The Affiliated Hospital, Xi'an Medical University , Xi'an , China.
authors.forename: Xinna
authors.initials: X
authors.lastname: Wang
authors.forename: Jianmei
authors.initials: J
authors.lastname: Luo
authors.forename: Albert Wingnang
authors.initials: AW
authors.lastname: Leung
authors.forename: Yajun
authors.initials: Y
authors.lastname: Li
authors.forename: Hongwei
authors.initials: H
authors.lastname: Zhang
authors.forename: Chuanshan
authors.initials: C
authors.lastname: Xu
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D002470:Cell Survival; D056945:Hep G2 Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008113:Liver Neoplasms / Q000473:pathology*; D053078:Membrane Potential, Mitochondrial; D010569:Perylene / Q000031:analogs & derivatives* / Q000378:metabolism; D021381:Protein Transport; D011809:Quinones / Q000378:metabolism*; D014464:Ultrasonic Therapy / Q000009:adverse effects*
chemical_list: D011809:Quinones; D010569:Perylene; C064522:hypocrellin B
keywords: Sonodynamic therapy; apoptosis; clonogenic survival; hepatocellular carcinoma cells; hypocrellin B; mitochondrial damage; subcellular localization