目的 通過生物信息學方法研究與鐵死亡相關的治療膽管癌潛在作用機制，高通量篩選通過鐵死亡途徑治療膽管癌疾病的潛在小分子藥物。方法 在GEO（Gene Expression Omnibus）在線數(shù)據(jù)庫中檢索與膽管癌相關的數(shù)據(jù)集，使用TCGA（The Cancer Genome Atlas）數(shù)據(jù)庫獲得膽管癌相關樣本及正常樣本的數(shù)據(jù)，將上述篩選得到的數(shù)據(jù)集矩陣進行規(guī)范化數(shù)據(jù)整理，通過Sanger平臺對數(shù)據(jù)集中的靶點基因進行l(wèi)imma差異分析，通過FerrDb鐵死亡相關分析數(shù)據(jù)庫收集與鐵死亡相關的靶點，將收集得到的鐵死亡相關靶點與差異分析得到的膽管癌相關疾病靶點取交集后，得到共同靶點，將上述共同靶點導入STRING數(shù)據(jù)庫，得到與鐵死亡相關的膽管癌疾病靶點間的蛋白質相互作用（PPI）網絡分析，篩選核心靶點，分別使用DAVID v6.8與Funrich富集分析軟件對上述潛在作用靶點進行生物學功能與信號通路的富集分析，并利用cMAP（Connectivity Map）數(shù)據(jù)庫高通量篩選通過鐵死亡途徑治療膽管癌疾病的潛在小分子藥物，將小分子化合物與靶蛋白進行分子對接分析，通過TIMER（Tumor Immune Estimation Resource）數(shù)據(jù)庫分析鐵死亡與膽管癌疾病共同靶點各自間表達水平與免疫細胞浸潤水平或免疫檢查點表達水平的相關性。結果 篩選得到3 133個膽管癌相關靶點，鐵死亡相關靶點487個，取交集處理共得到93個鐵死亡與膽管癌疾病相關的共同靶點，共同靶點的基因本體（GO）功能富集分析結果提示鐵死亡途徑對膽管癌治療方面可能與轉錄調控相關過程、凋亡與細胞增殖等密切相關，京都基因與基因組百科全書（KEGG）信號通路富集分析結果表明鐵死亡途徑對膽管癌治療方面可能與TRAJL分子信號通路、表皮生長因子受體（EGFR/ErbB）相關信號通路、鞘氨醇-1-磷酸受體-1（S1P1）分子信號通路、胰島素相關信號通路等過程密切相關，通過篩選得到3個小分子化合物（海恩酮、氯可托龍戊酸酯、利扎曲普坦），分子對接結果表明氯可托龍戊酸酯與其他2種成分相比，對上述3個潛在核心共同作用靶點（TP53、PTEN、SRC）的結合普遍較其余小分子成分強，從而提示其有望通過鐵死亡途徑治療膽管癌相關疾病。結論 以鐵死亡途徑作為切入點，通過生物信息學方法探尋治療膽管癌的分子作用機制及蛋白靶點，從而篩選得到潛在的治療藥物，具有重要意義。

Objective To screen the potential therapeutic targets of cholangiocarcinoma associated with ferroptosis using bioinformatics methods, and to high-throughput screen the potential small molecule drugs for the treatment of cholangiocarcinoma diseases through ferroptosis mechanisms. Methods Data sets related to cholangiocarcinoma were retrieved from GEO (Gene Expression Omnibus) online database, and the data of cholangiocarcinoma related samples and normal samples were obtained from TCGA (The Cancer Genome Atlas) database. The data set matrix obtained from the above screening was standardized, and limma difference analysis was performed on target genes in the data set through Sanger platform, and targets related to ferroptosis were collected through FerrDb ferroptosis related online database, and after intersecting the collected ferroptosis related targets with cholangiocarcinoma targets obtained through differential analysis methods, a common target is obtained. The common targets are imported into the STRING database to obtain protein-protein interaction (PPI) network analysis between ferroptosis related cholangiocarcinoma targets. Core targets are screened, and biological functional and signaling pathway enrichment analysis are performed on the above potential targets using DAVID v6.8 and Funrich enrichment analysis software. Using the cMAP (Connectivity MAP) database to high-throughput screen potential small molecule drugs for the treatment of cholangiocarcinoma through the ferroptosis pathway, and molecular docking analysis was performed on small molecule compounds and target proteins. The TIMER (Tumor Immune Estimation Resource) database was used to analyze the correlation between the expression levels of common targets of ferroptosis and cholangiocarcinoma, immune cell infiltration levels, or immune checkpoint expression levels. Results 3 133 targets related to cholangiocarcinoma and 487 targets related to ferroptosis were identified, and a total of 93 common targets related to ferroptosis and cholangiocarcinoma were obtained by intersection. The GO functional enrichment analysis of the common targets suggested that the ferroptosis pathway may be closely related to the treatment of cholangiocarcinoma. The KEGG signaling pathway enrichment analysis results showed that the ferroptosis pathway may be related to the TRAJL molecular signaling pathway in the treatment of cholangiocarcinoma. The EGF(ErbB) receptor related signaling pathway, S1P1 molecular signaling pathway, insulin related signaling pathway, and other processes are also closely related it. Meanwhile, some small molecule compounds were identified through screening, and a total of three components were identified. Molecular docking results showed that compared with the other two components, the binding of chlorotolone valerate to the three potential core targets mentioned above is generally stronger than the other small molecule components, this suggests that it has the potential to treat cholangiocarcinoma related diseases through the ferroptosis pathway. Conclusion Using the ferroptosis pathway as a starting point, potential targets for cholangiocarcinoma can be identified through bioinformatics methods, and potential therapeutic drugs can be screened.

R979.1

山東省藥學會醫(yī)院藥學科研專項項目（yyyx2021ms-04）