Atractylodin and β-eudesmol from Atractylodes lancea (Thunb.) DC. Inhibit Cholangiocarcinoma Cell Proliferation by Downregulating the Notch Signaling Pathway

Abstract

Objective: 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. Methods: 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. Results: Atractylodin and β-eudesmol moderately inhibited HuCCT-1 cell growth with IC 50 (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. Conclusion:The notch signaling pathway could be a promising target of atractylodin and β-eudesmol in CCA.

Liensinine inhibits progression of intrahepatic cholangiocarcinoma by regulating TGF‐β1 /P‐smad3 signaling through HIF‐1a

“…To further investigate the efficacy of liensinine in treatment of ICC, we tried to combine liensinine with gemcitabine. Based on existing studies, 43–45 we set the concentration of gemcitabine on RBE and Hucc‐T1 to 10μM. Through colony formation test, wound healing assays, migration and invasion Transwell experiments, we found that, in RBE and Hucc‐T1 cells, liensinine combined with gemcitabine had certain curative effects on the proliferation, as well as on migration and invasion (Figures ).…”

“…Based on existing studies, [43][44][45] tumor resistance to gemcitabine, [46][47][48][49] and therapy targeting on HIF-1a may improve the efficacy of gemcitabine. 50,51 Besides, available data does not rule out the possibility that gemcitabine directly or indirectly acts on HIF-1a.…”

Phytochemistry and Pharmacology of Sesquiterpenoids from Atractylodes DC. Genus Rhizomes

“…In addition, atractylon regulates the expression of thymopoietin antisense transcript 1 (TMPO-AS1) and coiled-coil domain-containing 183 antisense RNA 1 (CCDC183-AS1) and inhibits the invasion and migration of liver cancer cells [ 81 ]. β -eudesmol was found to have moderate activity against human cholangiocarcinoma (HuCCT-1) cell growth with an IC 50 (concentration that inhibits cell growth by 50%) value of 16.80 ± 4.41 µg/mL through the Notch signaling pathway and its upstream/downstream molecules in the CCA cell line at the gene and protein expression levels [ 82 ]. Moreover, β -eudesmol treatment (2.5–5 mg/kg) significantly inhibited the growth of H 22 and S 180 mouse tumor in vivo, which indicated that it inhibited angiogenesis via suppressing CREB activation in growth factor signaling pathway [ 83 ].…”

“…Eudesm-4(15)-ene-7α,11-diol (72), (5R,10S)-eudesm-4(15),7-diene-11-ol-9-one (73), and eudesm-4(15),7(11)-diene-9α,11-diol (74) were separated from A. lancea via silica gel column chromatography and preparative TLC [58]. Two new nitrogen-containing sesquiterpenoids, atractylenolactam A (75) and atractylenolactam B (76); two new sesquiterpene lactones, 8-methoxy-AT-V (77) and 15-acetoxyl AT-III (86); and four known analogs (78)(79)(80)(81)(82) were separated from A. macrocephala using column chromatography and preparative HPLC, and the absolute configurations were established using time-dependent density functional theory ECD (TDDFT-ECD) calculations [59,60]. Zhou et al [61] isolated six eudesmane-type sesquiterpenoids (83)(84)(85)(86)(87)(88) from A. lancea with repeated silica gel column chromatography, and their structures were determined using physiochemical and spectroscopic evidence.…”