title: Synergistic complex from plants Solanaceae exhibits cytotoxicity for the human hepatocellular carcinoma cell line HepG2.
abstract: BACKGROUND
  It had been demonstrated that sugars from various plants can act as potent agents, which induce apoptosis of cancer cells.
  
  
  METHODS
  Using HPLC, we fractionated a mixture of two plant extracts from the plant family Solanaceae, namely Capsicum chinense and the plant family Amaryllidaceae namely Allium sativum. We evaluated the effect of different fractions on apoptosis of HepG2 cell line. The most effective fraction was further studied to determine its molecular composition using mass spectrometry (MS) and NMR. We further evaluated the effect of determined molecular composition found in the selected fraction by using a mixture of commercially available substances, which were found in the fraction and tested its pro-apoptotic effect on HepG2 cells. To get some insight into potential apoptotic mechanisms we studied caspase-3 activity and mitochondrial integrity in treated cells.
  
  
  RESULTS
  Out of 93 fractions obtained by HPLC from the plant extract we found HPLC fraction 10 (10 min elution) was the most effective. MS and NMR studies revealed high presence of cellobiose together with vitamin C, sulphur (S) and trace amounts of selenium (Se). HPLC fraction 10 triggered apoptosis of HepG2 within 3 h in the 0.01-1.0 mg/mL concentration range. Furthermore, a mixture of pure cellobiose, vitamin C, S and Se (complex cellobiose/C/S/Se) had a very similar capacity in inducing apoptosis of HepG2 cells compared to HPLC fraction 10. Complex cellobiose/C/S/Se was capable of inducing caspase-3 activity and led to loss of mitochondrial integrity. The capacity of cellobiose alone to induce apoptosis of HepG2 was approximately 1000-fold lower compared to complex cellobiose/C/S/Se.
  
  
  CONCLUSIONS
  In this study we present the highly synergistic effect of a unique complex consisting of cellobiose, vitamin C, sulphur and selenium on triggering the apoptosis of human hepatocellular carcinoma (HepG2) cell line.
authors.affiliation: University of Ljubljana, Medical Faculty, Vrazov trg 2, SI-1000, Ljubljana, Slovenia. romina.schwarzlin@medisanus.com.
authors.forename: Romina
authors.initials: R
authors.lastname: Schwarzlin
authors.affiliation: University of Ljubljana, Medical Faculty, Vrazov trg 2, SI-1000, Ljubljana, Slovenia.
authors.forename: Nika
authors.initials: N
authors.lastname: Pušenjak
authors.affiliation: National Institute of Chemistry, Hajdrihova 19, SI-1000, Ljubljana, Slovenia.|EN-FIST Centre of Excellence, Dunajska 156, SI-1000, Ljubljana, Slovenia.
authors.forename: Damjan
authors.initials: D
authors.lastname: Makuc
authors.affiliation: National Institute of Chemistry, Hajdrihova 19, SI-1000, Ljubljana, Slovenia.
authors.forename: Mitja
authors.initials: M
authors.lastname: Križman
authors.affiliation: National Institute of Chemistry, Hajdrihova 19, SI-1000, Ljubljana, Slovenia.|EN-FIST Centre of Excellence, Dunajska 156, SI-1000, Ljubljana, Slovenia.
authors.forename: Irena
authors.initials: I
authors.lastname: Vovk
authors.affiliation: National Institute of Chemistry, Hajdrihova 19, SI-1000, Ljubljana, Slovenia.|EN-FIST Centre of Excellence, Dunajska 156, SI-1000, Ljubljana, Slovenia.
authors.forename: Janez
authors.initials: J
authors.lastname: Plavec
authors.affiliation: Centre for Development of Advanced Therapy Medicinal Products, Blood Transfusion Centre of Slovenia, Šlajmerjeva 6, SI-1000, Ljubljana, Slovenia. urban.svajger@ztm.si.
authors.forename: Urban
authors.initials: U
authors.lastname: Švajger
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular; D002470:Cell Survival / Q000187:drug effects*; D004357:Drug Synergism; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D019657:Solanaceae / Q000737:chemistry*
chemical_list: D000970:Antineoplastic Agents; D010936:Plant Extracts