atractylodin and Bile-Duct-Neoplasms

Notch signaling pathway has been reported to be involved in the development and progression of various types of cancer, including cholangiocarcinoma (CCA).  Compounds that modulate this signaling pathway could be promising candidates for CCA treatment and control. The study investigated the antiproliferative activities and modulatory effects of atractylodin and β-eudesmol, the two bioactive compounds of Atractylodes lancea (Thunb.) DC. , on Notch signaling and upstream molecules (Notch1 and Notch2 receptors, JAG1, mTOR, PI3K, and YAP), and downstream molecules (Snail) in HuCCT-1 (CCA cell line) and OUMS-36T-1 (normal fibroblast cell line). Gemcitabine (standard drug for CCA), and Notch inhibitors (DAPT and zebularine) were included in the experiments as positive control compounds.. The antiproliferative activity was evaluated using MTT assay.  mRNA and protein expression of Notch signaling molecules were evaluated using real-time PCR and Western blot analysis.. Atractylodin and β-eudesmol moderately inhibited HuCCT-1  cell growth with IC50 (concentration that inhibits cell growth by 50%) of 29.00 ± 6.44 and 16.80 ± 4.41 µg/ml (mean±SD), respectively. The direction and extent of the modulatory effects on mRNA and protein expression in the CCA cell line varied with the signaling molecules. Notch1 receptor was shown to be the most promising target of atractylodin and β-eudesmol in CCA. The level of gene expression was significantly downregulated (0.042 to 0.195 fold of control) after treating HuCC-T1 cells with both compounds at low and high concentrations. The extent and change in Notch1 gene expression correlated well with protein expression.. The notch signaling pathway could be a promising target of atractylodin and β-eudesmol in CCA.  
.

Topics: Atractylodes; Bile Duct Neoplasms; Bile Ducts, Intrahepatic; Cell Proliferation; Cholangiocarcinoma; Humans; Signal Transduction

The effects of atractylodin (ATD), the bioactive compound from Atractylodes lancea, on migration and autophagy status of cholangiocarcinoma cell line were investigated.. Cytotoxic activity and effects on cell migration and invasion were evaluated by MTT and trans-well assay, respectively. Autophagy and underlying molecular mechanisms were investigated using flow cytometry and western blot analysis.. ATD regulated the activity of PI3K/AKT/mTOR and p38MAPK signalling pathways which contributed to autophagy induction. HuCCT-1 cell growth was inhibited by ATD in a time- and dose-dependent manner. ATD inhibited the migration and invasion of HuCCT1 cells in a concentration-dependent manner. It also induced autophagy in HuCCT1 cells in a time- and dose-dependent manner. The SB202190 (autophagy inducer) and 3-MA (autophagy inhibitor) significantly increased and decreased the rate of ATD-induced autophagy, respectively. The 24 h exposure of ATD inhibited the phosphorylation of phosphatidylinositol-3-kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR), mitogen-activated protein kinase (p38MAPK) and increased Beclin-1 expression and LC3 conversion. It also reduced p-AKT/AKT, p-mTOR/mTOR and p-p38MAPK/p38MAPK.. ATD inhibits the proliferation and induces CCA cell autophagy via regulating PI3K/AKT/mTOR and p38MAPK signalling pathways.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Atractylodes; Autophagy; Beclin-1; Bile Duct Neoplasms; Bile Ducts, Intrahepatic; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cholangiocarcinoma; Furans; Humans; MAP Kinase Signaling System; Microtubule-Associated Proteins; p38 Mitogen-Activated Protein Kinases; Phosphatidylinositol 3-Kinase; Phosphatidylinositol 3-Kinases; Phytotherapy; Plant Extracts; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases

The study aimed to identify potential cell signaling pathways and protein targets of actions of atractylodin and β-eudesmol in cholangiocarcinoma, the two active compounds isolated from Atracylodes lancea using proteomics approach.. The cholangiocarcinoma cell line, CL-6, was treated with each compound for 3 and 6 hours, and the proteins from both intra- and extracellular components were extracted. LC-MS/MS was applied following the separation of the extract proteins by SDS-PAGE and digestion with trypsin. Signaling pathways and protein expression were analyzed by MASCOT and STITCH software.. A total of 4,323 and 4,318 proteins were identified from intra- and extracellular components, respectively. Six and 4 intracellular proteins were linked with the signaling pathways (apoptosis, cell cycle control, and PI3K-AKT) of atractylodin and β-eudesmol, respectively. Four and 3 extracellular proteins were linked with the signaling pathways (NF-κB and PI3K-AKT) of atractylodin and β-eudesmol, respectively.. In conclusion, a total of 17 proteins associated with four cell signaling pathways that could be potential molecular targets of anticholangiocarcinoma action of atractylodin and β-eudesmol were identified through the application of proteomics approach.

Topics: Apoptosis; Bile Duct Neoplasms; Cell Cycle; Cell Line, Tumor; Cholangiocarcinoma; Furans; Humans; Proteomics; Sesquiterpenes, Eudesmane; Signal Transduction

The anti-cholangiocarcinoma (CCA) activity of atractylodin isolated from Atractylodes lacea (Thunb.) DC. has previously been demonstrated both in vitro and in vivo. However, the compound is insoluble in water and must be dissolved in organic solvent which might be harmful to human body. The aim of the study was to develop atractylodin-loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) (ALNPs) and to investigate its cytotoxic activity against CCA.. The ALNPs were prepared using PLGA MW 12,000 and 48,000 by solvent displacement methods. Particle size, polydispersity index (PDI), zeta potential, encapsulation efficiency (%EE) and loading efficiency (%LE) as well as drug releasing profile of ALNPs were characterized. The selected ALNPs formulation was then investigated cytotoxic activity against CCA cell lines, CL-6 and HuCC-T1.. The ALNPs preparation was achieved using PLGA MW 12,000 (ALNPs-1) with mean (±SD) values of particle diameter, PDI and zeta potential of 158.13±0.21 nm, 0.076±0.003, and (-) 23.80± (-) 0.75 mV, respectively. The transmission electron microscopy (TEM) showed spherical morphology of NPs. The %EE and %LE were 50.16±1.77% and 2.22±0.08%, respectively. The release of atractylodin from ALNPs-1 in PBS was up to 88% in 72 h. The potency of ALNPs-1 cytotoxic activity including selectivity against CCA cell line, CL-6, were about twice of the unformulated atractylodin after 24 h of exposure (IC50: 29.28 vs 56.36 µg/mL, selectivity index 2.99 vs 1.50).. ALNPs were successfully prepared by solvent displacement method using PLGA MW 12,000 (ALNPs-1) with suitable pharmaceutical properties and cytotoxic activity against CCA. However, nano-formulation with improved pharmaceutical properties (higher %EE and %LE) and cytotoxic activity (improved selectivity to CCA) should be further developed for potential used as drug delivery systems for the treatment of CCA.
.

Topics: Antineoplastic Agents; Bile Duct Neoplasms; Cholangiocarcinoma; Drug Carriers; Drug Delivery Systems; Furans; Humans; Nanoparticles; Polylactic Acid-Polyglycolic Acid Copolymer; Tumor Cells, Cultured

To evaluate the cytotoxic activity of atractylodin and its potential effects on heme oxygenase (HO)-1 production, STAT1/3 phosporylation and major NF-κB protein expression in the cholangiocarcinoma-associated cell line CL-6.. Standard MTT assay was used for accessing antiproliferative activity on CL-6 cells. Normal human embryonic fibroblast (OUMS) cell was taken as control cell line. Colony formation and wound healing assay were conducted to access the effects of atractylodin on cell proliferation and directional migration activity of CL-6 cells. Western blot was used for evaluating levels of protein expression and phosphorylation.. Atractylodin exerts significant cytotoxic activity against CL-6 cells which may be linked to its suppressive effect on HO-1 production, STAT1/3 phosphorylation and expression of key NF-κB proteins.

Topics: Antineoplastic Agents, Phytogenic; Bile Duct Neoplasms; Blotting, Western; Cell Line; Cell Line, Tumor; Cell Proliferation; Cholangiocarcinoma; Dose-Response Relationship, Drug; Furans; Heme Oxygenase-1; Humans; Inhibitory Concentration 50; NF-kappa B; Phosphorylation; STAT1 Transcription Factor; STAT3 Transcription Factor; Time Factors

Topics: Antineoplastic Agents; Apoptosis; Bile Duct Neoplasms; Caspase 3; Caspase 7; Cell Cycle Checkpoints; Cell Line, Tumor; Cholangiocarcinoma; Dose-Response Relationship, Drug; Furans; G1 Phase; Humans; Sesquiterpenes, Eudesmane; Time Factors