Suppression of Cholangiocarcinoma Cell Growth and Proliferation by Atractylodes lancea (Thunb) DC. through ERK-Signaling Cascade

Abstract

Objective:The study aimed to investigate the inhibitory effects of AL on the ERK signaling molecules (ERK, p-ERK, cyclin D, and eIF4B) and the growth and proliferation of CCA cells. Materials and methods: The viability of the three CCA cell lines CL-6, HuCCT1, and HuH28 was determined using MTT assay. The effect of Ras/ERK inhibitors on protein expression in the presence of AL extract was investigated. The protein extracted from each CCA cell following exposure to AL and/or Ras/ERK inhibitors were separated on 12.5% SDS-PAGE. The analysis of mRNA expression following 48 and 72 hours of AL exposure in comparison with 0 hours (non-exposed cells) was performed by using RT-PCR. Results: The potency of cytotoxic activity of AL (by MTT assay) was about three times higher than the standard drug 5-fluorouracil. The IC 50 (concentration that inhibits cell growth by 50%) of AL for the CL-6, HuCCT-1 and HuH28 cell lines were 29.77±6.64, 35.45±4.96, and 35.32±6.69 µg/mL (mean+SD), respectively. The cells were exposed to AL extract at the IC 50 for 0, 12, 24, 48, and 72 hours in the absence and presence of Ras/ERK inhibitors (salirasib and XMD8-92). Protein expression was determined by Western blot analysis. The results suggested the lack of significant inhibitory effect of AL on ERK at 48 and 72 hours of exposure in all CCA cell types. On the other hand, a significant inhibitory effect was observed with p-ERK expression in all CCA cell types. Cyclin D was significantly down-regulated at 72 hours of exposure in all cell types with different potencies. The expression of eIF4B was markedly inhibited in HuCCT-1 but slightly inhibited in CL-6 and HuH28 cells. Real-time PCR analysis revealed significant down-regulation of ERK following 72 hours of AL exposure in the HuCCT1 and HuH28, but not CL-6 cell. Conclusion: The ERK signaling cascade and downstream molecules are potential targets of action of AL in CCA.

Anti-Inflammatory Effect of Garcinol Extracted from Garcinia dulcis via Modulating NF-κB Signaling Pathway

“…Cell viabilities of THP-1 and RAW 264.7 cells were measured by using a 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) assay [ 33 , 34 ]. Briefly, 1 × 10 4 cells per well were plated into a 96-well microtiter plate using a complete culture medium and incubated for 24 h. After incubation, the culture medium was removed.…”

“…For determining the effects of garcinol, the co-incubation of LPS and garcinol (10, 20, and 30 µM) or 5 µM of dexamethasone (Sigma Aldrich, Darmstadt, Germany) as a positive control anti-inflammatory agent were performed. In addition, the half-maximal inhibitory concentration (IC 50 ) was evaluated as previously described [ 33 , 34 ].…”

Ethyl Acetate Extract of Halymenia durvillei Induced Apoptosis, Autophagy, and Cell Cycle Arrest in Colorectal Cancer Cells

“…The gene expression of PI3K, AKT, mTOR, and glyceraldehyde 3-phosphate dehydrogenase (a housekeeping control) in HDEA-treated HT-29 cells was evaluated. The qRT-PCR was performed following the previously described procedure (Martviset et al, 2021). The total RNA was extracted, and the first-strand cDNA from the purified total RNA was synthesized.…”

“…In traditional East Asian medicine, seaweed has been used as a nutraceutical to diminish cancer severity (Brown et al, 2014). Previous studies revealed several mechanisms of seaweed extracts that exert anticancer effects (Farooqi et al, 2012;Chantree et al, 2021). In CRC, there is evidence suggesting the anticancer activities of seaweed (Alves et al, 2018;Deviyani-Zakaria et al, 2018;Al-Aadily et al, 2022).…”

Modulatory Effects of Atractylodin and β-Eudesmol on Human Cytochrome P450 Enzymes: Potential Drug-Drug Interactions

“…Both are the primary active ingredients responsible for the majority of the pharmacologic properties of AL [ 6 ]. Recently, promising activities against cholangiocarcinoma of atractylodin and β-eudesmol have been demonstrated in a series of non-clinical studies [ 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 ]. With regard to metabolic drug interactions, the crude ethanolic extract of AL rhizomes was shown to potently inhibit human CYP1A2 in vitro, with comparable potency with the reference hCYP1A2 inhibitor—α-naphthoflavone.…”