目的 采用網(wǎng)絡藥理學結合實驗驗證方法探索去甲漢黃芩素對口腔鱗狀細胞癌的抗腫瘤活性及靶點。方法 采用網(wǎng)絡藥理學方法分析去甲漢黃芩素及口腔鱗狀細胞癌的靶點。使用STRING和Cytoscape軟件構建蛋白相互作用（PPI）網(wǎng)絡?；虮倔w（GO）和京都基因和基因組百科全書（KEGG）富集分析生物學功能。采用分子對接和CETSA驗證去甲漢黃芩素與其關鍵靶標之間的結合能力。MTT和吖啶橙/溴化乙錠（AO/EB）法檢測去甲漢黃芩素對口腔鱗狀細胞癌細胞增殖以及凋亡的影響。免疫蛋白印跡法檢測相關蛋白的表達。結果 去甲漢黃芩素與口腔鱗狀細胞癌共有71個重疊靶點；分別涉及磷脂酰肌醇3-激酶（PI3K）/蛋白激酶（Akt）、氧化應激以及凋亡通路中相關基因；其中前5位的Hub基因為表皮生長因子受體（EGFR）、雌激素受體1（ESR1）、非受體酪氨酸激酶（SRC）、前列腺素內過氧化物酶2（PTGS2）、基質金屬蛋白酶9（MMP9）。分子對接結果顯示，去甲漢黃芩素與EGFR的結合力最強（-6.6 kcal/mol）；CETSA實驗和Kaplan-Meier生存曲線結果顯示，EGFR對口腔鱗狀細胞癌患者生存影響最大。實驗驗證結果顯示，去甲漢黃芩素以劑量相關性性地降低口腔鱗狀細胞癌細胞活性，IC₅₀為15 μmol/L。AO/EB染色分析顯示，去甲漢黃芩素誘導口腔鱗狀細胞癌細胞發(fā)生程序性凋亡來抑制癌細胞的活力，且與Bax水平的增加和Bcl-2水平的降低相關。結論 去甲漢黃芩素可能通過調控EGFR的表達進而影響PI3K/Akt、氧化應激以及凋亡途徑誘導口腔鱗狀細胞癌細胞凋亡。

Objective To investigate the antitumor activity and target of norwogonin in oral squamous cell carcinoma by network pharmacology combined with experimental verification. Methods The target of norwogonin and oral squamous cell carcinoma was analyzed by network pharmacology. Build a protein interaction (PPI) network using STRING and Cytoscape software. GO and KEGG enrichment analyze biological functions. Molecular docking and CETSA were used to verify the binding ability between norwogonin and its key targets. MTT and acridine orange/ethidium bromide (AO/EB) methods were used to detect the effects of norwogonin on the proliferation and apoptosis of oral squamous cell carcinoma cells. The expression of related proteins was detected by Immunowestern blotting. Results A total of 71 overlapping targets between norwogonin and oral squamous cell carcinoma, implicated in the PI3K/Akt, oxidative stress, and apoptosis signaling pathways, respectively. The top five Hub genes identified in this study are EGFR, ESR1, SRC, PTGS2, and MMP9. The molecular docking results showed that the binding force between norwogonin and EGFR was the strongest (-6.6 kcal/mol). The results of CETSA test and Kaplan-Meier survival curve showed that EGFR had the greatest impact on the survival of patients with oral squamous cell carcinoma. The experimental results showed that the activity of oral squamous cell carcinoma was decreased dose-dependent with IC₅₀ of 15 μmol/L. AO/EB staining analysis showed that norwogonin induced programmed apoptosis of oral squamous cell cancer cells to inhibit cancer cell viability, which was associated with increased Bax levels and decreased Bcl-2 levels. Conclusions Norwogonin may affect PI3K/Akt, oxidative stress and apoptosis pathway to induce apoptosis of oral squamous cell carcinoma cells by regulating EGFR expression.

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