| 引用本文: | 徐佳禛,吴加元,王涛,张慧,姜宁华.基于细胞实验及网络药理学分析桂枝改善非小细胞肺癌耐药的分子机制[J].中国现代应用药学,2025,42(1):1-8. |
| Xu Jiazhen,Wu Jiayuan,Wang Tao,Zhang Hui,Jiang Ninghua.A molecular mechanism analysis of Ramulus Cinnamomi in improving drug resistance of non-small cell lung cancer (NSCLC) using cell experiments and network pharmacology[J].Chin J Mod Appl Pharm(中国现代应用药学),2025,42(1):1-8. |
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| 基于细胞实验及网络药理学分析桂枝改善非小细胞肺癌耐药的分子机制 |
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徐佳禛1, 吴加元1, 王涛2,3, 张慧1, 姜宁华1
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1.嘉兴市第二医院;2.浙江省数字医学诊断技术重点实验室;3.杭州迪安医学检验中心有限公司
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| 摘要: |
| 目的 研究中药桂枝对非小细胞肺癌(NSCLC)耐药细胞株H1650增殖、转移、凋亡的影响,揭示桂枝改善H1650吉非替尼耐药性的功能及潜在分子机制。方法 在H1650中,使用桂枝、吉非替尼单药以及桂枝与吉非替尼联合用药,利用甲基偶氮唑盐比色法(MTT法)、细胞划痕法、流式细胞术,分别检测H1650细胞的增殖、迁移、凋亡情况。随后,利用网络药理学预测桂枝的有效活性成分及其影响NSCLC细胞功能的可能分子靶点,构建“桂枝-活性成分-靶点-疾病”的关系网络,并对桂枝潜在作用分子靶点进行Gene Oncology(GO)功能分析和Kyoto Encyclopedia of Genes and Genomes(KEGG)信号通路富集分析,预测与桂枝活性成分密切相关的核心分子及信号通路。最后,我们利用Western blot实验检测预测得到的核心分子蛋白质表达及蛋白质磷酸化修饰水平在桂枝、吉非替尼作用后的变化情况。结果 桂枝单药可抑制H1650细胞增殖、迁移,促进细胞凋亡；与吉非替尼单药组相比,桂枝联合吉非替尼可降低吉非替尼抑制H1650细胞增殖的IC50。网络药理学预测得到7个桂枝活性成分、252个NSCLC疾病相关分子靶点及多条与肿瘤发生发展密切相关的信号通路。其中,Src, PI3K, MAPK1及其相关信号通路可能是桂枝影响NCSLC细胞功能及耐药性的核心分子靶点。分子水平上,桂枝的确降低了H1650细胞Src蛋白质表达水平以及Src信号通路下游分子——Akt和mTOR的磷酸化水平。结论 桂枝可通过抑制Src/Akt/mTOR信号通路,改善H1650细胞对吉非替尼的耐药现象。 |
| 关键词: 桂枝 非小细胞肺癌 网络药理学 EGFR-TKIs Src |
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| A molecular mechanism analysis of Ramulus Cinnamomi in improving drug resistance of non-small cell lung cancer (NSCLC) using cell experiments and network pharmacology |
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Xu Jiazhen1, Wu Jiayuan1, Wang Tao2,3, Zhang Hui1, Jiang Ninghua1
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1.The Second Affiliated Hospital of Jiaxing University;2.Key Laboratory of Clinical in Vitro Diagnostic Techniques of Zhejiang Province;3.Dian Diagnostics Group CO., LTD.
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| Abstract: |
| ABSTRACT: OBJECTIVE To investigate the effect and potential molecular mechanism of Ramulus Cinnamomi on the proliferation, metastasis, and apoptosis of the drug-resistant non-small cell lung cancer (NSCLC) cell line H1650. METHODS H1650 cells were treated with Ramulus Cinnamomi extract, gefitinib, or two in combination. Using tetrazolium blue (MTT), wound healing assay, and flow cytometry, we studied each group's effect on cell viability, migration, and apoptosis. Then, network pharmacology was used to predict the effective active components of Ramulus Cinnamomi extract and the possible molecular targets relating to NSCLC, and to conduct the relationship network of "Ramulus Cinnamomi - active ingredient - targets - disease". GO and KEGG analysis was subsequently performed on the potential molecular targets of Ramulus Cinnamomi, and the core targets and signaling pathways that are closely related to the active components of Ramulus Cinnamomi were further predicted. Finally, Western blot was used to detect the changes in protein expression and protein phosphorylation level of the predicted core molecules after treatment with Ramulus Cinnamomi extract and gefitinib. RESULTS In H1650, Ramulus Cinnamomi extract inhibits cell viability and migration and prompts cell apoptosis. Compared to the single-drug group either with Ramulus Cinnamomi extract or gefitinib, those two drugs in combination reduce the IC50 of gefitinib inhibiting cell proliferation. Network pharmacology predicts 7 active components, 252 molecular targets related to NSCLC disease, and a number of signaling pathways closely related to the initiation and development of tumors. In particular, Src, PI3K, MAPK1, and their related signaling pathways may be the core molecular targets affecting NCSLC cell function and drug resistance. At the molecular level, Ramulus Cinnamomi extract significantly reduces the protein expression of Src and the phosphorylation level of Akt and mTOR in H1650 cells. CONCLUSION Ramulus Cinnamomi improves gefitinib resistance in H1650 cells presumably by inhibiting Src/Akt/mTOR signaling pathway. |
| Key words: ramulus cinnamomi NSCLC network pharmacology EGFR-TKIs Src |
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