forbesione and Bile-Duct-Neoplasms

Background: Chemotherapy for advanced cholangiocarcinoma (CCA) is largely ineffective; thus innovative\ combinations of chemotherapeutic agents and natural compounds represent a promising strategy. This study aimed to\ investigate the synergistic effects of forbesione combined with 5-fluorouracil (5-FU) in hamster cholangiocarcinoma\ (Ham-1) cells both in vitro and in vivo. The anti-tumor effects of 5-FU combined with forbesione in vitro were determined\ using the Sulforhodamine B (SRB) assay and the effects in vivo were assessed in transplanted Ham-1 allograph\ models. Using ethidium bromide/acridine orange (EB/AO) staining, the morphological changes of apoptotic cells\ was investigated. The expressions of apoptosis-related molecules after combined treatment with forbesione and 5-FU\ were determined using real-time RT-PCR and western blot analysis. Forbesione or 5-FU alone inhibited proliferation\ of Ham-1 cells in a dose-dependent manner and their combination showed a synergistic proliferation inhibitory effect\ in vitro. In vivo studies, forbesione in combination with 5-FU exhibited greater inhibition of the tumor in the hamster\ model compared with treatment using either drug alone. Forbesione combined with 5-FU exerted stronger apoptotic\ induction in Ham-1 cells than did single drug treatment. The combination of drugs strongly suppressed the expression\ of B-cell lymphoma 2 (Bcl-2) and procaspase-3 while enhancing the expression of p53, Bcl-2-associated X protein\ (Bax), apoptotic protease activating factor-1 (Apaf-1), caspase-9 and caspase-3, compared with single drug treatments.\ These results explained the decreased expression of cytokeratin 19 (CK19) positive cells and proliferation cell nuclear\ antigen (PCNA) positive cells in Ham-1 cell tumor tissues of the treated hamsters. There was no apparent systemic\ toxicity observed in the treated animals compared with the control groups. Forbesione combined with 5-FU strongly\ induced apoptosis in Ham-1 cells. The growth inhibitory effect of combined treatment using these two drugs was much\ greater than treatment with either drug alone, both in vitro and in vivo.

Topics: Animals; Antimetabolites, Antineoplastic; Bile Duct Neoplasms; Cholangiocarcinoma; Cricetinae; Drug Synergism; Fluorouracil; Garcinia; Heterocyclic Compounds; Plant Extracts; Tumor Cells, Cultured

To investigate the growth inhibitory mechanism of four caged xanthones from Garcinia hanburyi in cholangiocarcinoma (CCA) KKU-100 and KKU-M156 cells.. Four caged xanthones, selected on the basis of their anticancer potency and chemical structure diversities (i.e. isomorellin, isomorellinol, forbesione and gambogic acid) were used in this study. Growth inhibition of these caged xanthones was determined using the sulforhodamine B assay. Induction of apoptosis was assessed by observing cell morphology, ethidium bromide and acridine orange staining and DNA fragmentation assay. Levels of apoptotic-related gene and protein expressions were determined by a real-time reverse transcriptase polymerase chain reaction and Western blotting analysis, respectively.. The compounds were found to inhibit growth of both cell lines in a dose-dependent manner and also showed selective cytotoxicity against the cancer cells when compared with normal peripheral blood mononuclear cells. Growth suppression by these compounds was due to apoptosis, as evidenced by the cell morphological changes, chromatin condensation, nuclear fragmentation, and DNA ladder formation. At the molecular level, these compounds induced down-regulation of Bcl-2 and survivin proteins with up-regulation of Bax and apoptosis-inducing factor proteins, leading to the activation of caspase-9 and -3 and DNA fragmentation. The functional group variations did not appear to affect the anticancer activity with regard to the two CCA cell lines; however, at a mechanistic level, isomorellinol exhibited the highest potency in increasing the Bax/Bcl-2 protein expression ratio (120 and 41.4 for KKU-100 and KKU-M156, respectively) and in decreasing survivin protein expression (0.01 fold as compared to control cells in both cell lines). Other activities at the molecular level indicate that functional groups on the prenyl side chain may be important.. Our findings for the first time demonstrate that four caged xanthones induce apoptosis in CCA cells which is mediated through a mitochondria-dependent signaling pathway.

Topics: Apoptosis; Base Sequence; Bile Duct Neoplasms; Bile Ducts, Intrahepatic; Cell Line, Tumor; Cell Proliferation; Cholangiocarcinoma; DNA Primers; Garcinia; Gene Expression; Heterocyclic Compounds; Humans; Xanthones