Graduate Program in Bioclinical Sciences, Chulabhorn International College of Medicine, Thammasat University, Paholyothin Road, Klonglung, Pathumthani 12120, Thailand.
Center of Excellence in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma, Thammasat University, Paholyothin Road, Klonglung, Pathumthani 12120, Thailand.
Asian Pac J Cancer Prev. 2023 Feb 1;24(2):551-558. doi: 10.31557/APJCP.2023.24.2.551.
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.
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已报道 Notch 信号通路参与多种类型癌症的发展和进展，包括胆管癌（CCA）。调节此信号通路的化合物可能是 CCA 治疗和控制的有前途的候选药物。本研究调查了苍术内酯和 β-桉叶醇这两种苍术（Thunb.）DC 的生物活性化合物对 Notch 信号及其上游分子（Notch1 和 Notch2 受体、JAG1、mTOR、PI3K 和 YAP）以及下游分子（Snail）的增殖活性和调节作用在 HuCCT-1（CCA 细胞系）和 OUMS-36T-1（正常成纤维细胞系）中。吉西他滨（CCA 的标准药物）和 Notch 抑制剂（DAPT 和 zebularine）被包括在实验中作为阳性对照化合物。
使用 MTT 测定法评估增殖活性。使用实时 PCR 和 Western blot 分析评估 Notch 信号分子的 mRNA 和蛋白表达。
苍术内酯和 β-桉叶醇适度抑制 HuCCT-1 细胞生长，IC50（抑制细胞生长 50%的浓度）分别为 29.00±6.44 和 16.80±4.41μg/ml（平均值±标准差）。苍术内酯和 β-桉叶醇对 CCA 细胞系中 mRNA 和蛋白表达的调节方向和程度因信号分子而异。Notch1 受体被证明是苍术内酯和 β-桉叶醇在 CCA 中最有前途的靶标。用两种化合物在低浓度和高浓度处理 HuCC-T1 细胞后，基因表达水平显著下调（对照的 0.042 至 0.195 倍）。Notch1 基因表达的程度和变化与蛋白表达密切相关。
Notch 信号通路可能是苍术内酯和 β-桉叶醇在 CCA 中的有前途的靶标。
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