PMID- 19437563 OWN - NLM STAT- MEDLINE DCOM- 20090713 LR - 20220328 IS - 2219-2840 (Electronic) IS - 1007-9327 (Print) IS - 1007-9327 (Linking) VI - 15 IP - 18 DP - 2009 May 14 TI - Induction of apoptosis in human liver carcinoma HepG2 cell line by 5-allyl-7-gen-difluoromethylenechrysin. PG - 2234-9 AB - AIM: To investigate the effect of 5-allyl-7-gen-difluoromethylenechrysin (ADFMChR) on apoptosis of human liver carcinoma HepG2 cell line and the molecular mechanisms involved. METHODS: HepG2 cells and L-02 cells were cultured in vitro and the inhibitory effect of ADFMChR on their proliferation was measured by MTT assay. The apoptosis of HepG2 cells was determined by flow cytometry (FCM) using propidium iodide (PI) fluorescence staining. DNA ladder bands were observed by DNA agarose gel electrophoresis. The influence of ADFMChR on the proxisome proliferator-activated receptor gamma (PPARgamma), NF-kappaB, Bcl-2 and Bax protein expression of HepG2 cells were analyzed by Western blotting. RESULTS: MTT assay showed that ADFMChR significantly inhibited proliferation of HepG2 cells in a dose-dependent manner, with little effect on growth of L-02 cells, and when IC(50) was measured as 8.45 micromol/L and 191.55 micromol/L respectively, the potency of ADFMChR to HepG2 cells, was found to be similar to 5-fluorouracil (5-FU, IC(50) was 9.27 micromol/L). The selective index of ADFMChR cytotoxicity to HepG2 cells was 22.67 (191.55/8.45), higher than 5-FU (SI was 7.05 (65.37/9.27). FCM with PI staining demonstrated that the apoptosis rates of HepG2 cells treated with 3.0, 10.0 and 30.0 micromol/L ADFMChR for 48 h were 5.79%, 9.29% and 37.8%, respectively, and were significantly higher when treated with 30.0 micromol/L ADFMChR than when treated with 30.0 micromol/L ChR (16.0%) (P < 0.05) and were similar to those obtained with 30.0 micromol/L 5-FU (41.0%). DNA agarose gel electrophoresis showed that treatment of HepG2 cells with 10.0 micromol/L ADFMChR for 48 h and 72 h resulted in typical DNA ladders which could be reversed by 10.00 micromol/L GW9662, a blocker of PPARgamma. Western blotting analysis revealed that after 24 h of treatment with 3.0, 10.0, 30.0 micromol/L ADFMChR, PPARgamma and Bax protein expression in HepG2 cells increased but Bcl-2 and NF-kappaB expression decreased; however, pre-incubation with 10.0 micromol/L GW9662 could efficiently antagonize and weaken the regulatory effect of 3.0, 30.0 micromol/L ADFMChR on PPARgamma and NF-kappaB protein expression in HepG2 cells. CONCLUSION: ADFMChR induces apoptosis of HepG2 cell lines by activating PPARgamma, inhibiting protein expression of Bcl-2 and NF-kappaB, and increasing Bax expression. FAU - Tan, Xiang-Wen AU - Tan XW AD - Laboratory of Medicine Engineering, Medical College, Hunan Normal University, Changsha 410006, Hunan Province, China. FAU - Xia, Hong AU - Xia H FAU - Xu, Jin-Hua AU - Xu JH FAU - Cao, Jian-Guo AU - Cao JG LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - World J Gastroenterol JT - World journal of gastroenterology JID - 100883448 RN - 0 (5-allyl-7-gen-difluoromethylenechrysin) RN - 0 (Flavonoids) RN - 0 (NF-kappa B) RN - 0 (PPAR gamma) RN - 0 (Proto-Oncogene Proteins c-bcl-2) RN - 0 (bcl-2-Associated X Protein) SB - IM MH - Animals MH - Apoptosis/*drug effects MH - *Carcinoma, Hepatocellular MH - *Cell Line, Tumor/drug effects/physiology MH - DNA Fragmentation MH - Flavonoids/*pharmacology MH - Humans MH - *Liver Neoplasms MH - NF-kappa B/metabolism MH - PPAR gamma/metabolism MH - Proto-Oncogene Proteins c-bcl-2/metabolism MH - bcl-2-Associated X Protein/metabolism PMC - PMC2682238 EDAT- 2009/05/14 09:00 MHDA- 2009/07/14 09:00 PMCR- 2009/05/14 CRDT- 2009/05/14 09:00 PHST- 2009/05/14 09:00 [entrez] PHST- 2009/05/14 09:00 [pubmed] PHST- 2009/07/14 09:00 [medline] PHST- 2009/05/14 00:00 [pmc-release] AID - 10.3748/wjg.15.2234 [doi] PST - ppublish SO - World J Gastroenterol. 2009 May 14;15(18):2234-9. doi: 10.3748/wjg.15.2234.