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蛋白质组学分析用于鉴定苍术素和β-桉叶醇抗胆管癌的潜在细胞信号通路及作用蛋白靶点

Proteomics Analysis for Identification of Potential Cell Signaling Pathways and Protein Targets of Actions of Atractylodin and β-Eudesmol Against Cholangiocarcinoma.

作者信息

Kotawong Kanawut, Chaijaroenkul Wanna, Roytrakul Sittiruk, Phaonakrop Narumon, Na-Bangchang Kesara

机构信息

Chulabhorn International College of Medicine, Thammasat University, Paholyothin Road, Klonglung, Pathumthani Thailand.

Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani Thailand.

出版信息

Asian Pac J Cancer Prev. 2020 Mar 1;21(3):621-628. doi: 10.31557/APJCP.2020.21.3.621.

Abstract

OBJECTIVE

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.

METHOD

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.

RESULTS

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.

CONCLUSION

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.

摘要

目的

本研究旨在利用蛋白质组学方法,确定茅术醇和β-桉叶醇(从苍术中分离出的两种活性化合物)在胆管癌中的潜在细胞信号通路和作用蛋白靶点。

方法

用每种化合物处理胆管癌细胞系CL-6 3小时和6小时,提取细胞内和细胞外成分的蛋白质。通过SDS-PAGE分离提取的蛋白质并用胰蛋白酶消化后,应用液相色谱-串联质谱法(LC-MS/MS)。通过MASCOT和STITCH软件分析信号通路和蛋白质表达。

结果

分别从细胞内和细胞外成分中鉴定出4323种和4318种蛋白质。分别有6种和4种细胞内蛋白质与茅术醇和β-桉叶醇的信号通路(凋亡、细胞周期调控和PI3K-AKT)相关联。分别有4种和3种细胞外蛋白质与茅术醇和β-桉叶醇的信号通路(NF-κB和PI3K-AKT)相关联。

结论

总之,通过应用蛋白质组学方法,共鉴定出17种与四种细胞信号通路相关的蛋白质,这些蛋白质可能是茅术醇和β-桉叶醇抗胆管癌作用的潜在分子靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb88/7437331/5b6e6e9a4a1c/APJCP-21-621-g001.jpg

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