Molecular mechanism of Salvia miltiorrhiza inhibiting FASN expression in the treatment of bladder cancer based on network pharmacology analysis

doi:10.3760/cma.j.issn.1007-1245.2023.23.022

Abstract

Abstract:

Objective　To study the active ingredients and potential molecular targets of Salvia miltiorrhiza and reveal the molecular mechanism of Salvia miltiorrhiza in the treatment of bladder cancer by using the ideas and methods of network pharmacology. Methods　The study was conducted from January 2022 to June 2023. The active ingredients in Salvia miltiorrhiza with oral bioavailability (OB) ≥30% and drug-likeness (DL) ≥0.18 were analyzed based on Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and then the downstream targets of the active ingredients were obtained for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) function enrichment analysis, which were included in the bladder cancer dataset in the Cancer Genome Atlas (TCGA) database for risk regression analysis, combined with the expression levels of risk genes and survival prognosis information to screen the potential therapeutic targets of Salvia miltiorrhiza in bladder cancer, and validation and in vitro treatment experiments were carried out in bladder cancer cell lines. Results　A total of 65 active ingredients in Salvia miltiorrhiza could act on 270 downstream target proteins, and its function enrichment analysis focused on nitrogen metabolism, bladder cancer, endocrine resistance, prostate cancer, and neuroactive ligand receptor interaction, which indicated that Salvia miltiorrhiza had the potential to treat bladder cancer. Sequencing data of bladder cancer patients showed that 44 genes had prognostic value in bladder cancer (P<0.05). The study further successfully constructed a risk regression model based on 18 genes among which [AUC (area under the curve)=0.819]. Combined with the expression levels of genes in cancer and adjacent cancer, MMP9, MAOA, PSMB5, HCAR and FASN were screened as molecular targets of Salvia miltiorrhiza for bladder cancer treatment, among which FASN had the highest risk coefficient (core=0.007 412). Cell experiment showed that the expression of FASN in bladder cancer cell line J82 was significantly increased, while after treatment with Salvia miltiorrhiza (IC50=10 mg/L), the proliferation, invasion, and migration of bladder cancer cells were significantly reduced, and the expression of FASN protein was also reduced. Conclusion　This study revealed the effective active ingredients of Salvia miltiorrhiza and its molecular targets for bladder cancer treatment, and provided the necessary experimental basis and theoretical basis for the clinical application of Salvia miltiorrhiza in the treatment of bladder cancer.

Key words:

Bladder cancer, Salvia miltiorrhiza, FASN, Network pharmacology

摘要：

目的　应用网络药理学思路与方法研究丹参的有效成分及潜在分子靶点，揭示丹参治疗膀胱癌的分子机制。方法　研究时间为2022年1月至2023年6月。基于中药系统药理学数据库分析平台（TCMSP）分析丹参中生物利用度（OB）≥30%，类药性指数（DL）≥0.18的活性成分，进而获得活性成分的下游靶点进行基因本体论（GO）与京都基因与基因组百科全书（KEGG）功能富集分析，并纳入癌症基因图谱（TCGA）数据库中膀胱癌数据集进行风险回归分析，结合风险基因表达水平与生存预后信息筛选丹参在膀胱癌中的潜在治疗靶点，并在膀胱癌细胞系中进行验证与体外治疗实验。结果　丹参中65个活性成分可作用于下游270个靶蛋白，其功能富集分析集中在氮代谢、膀胱癌、内分泌抵抗、前列腺癌、神经活性配体-受体相互作用等通路上，提示丹参具备治疗膀胱癌的潜力，膀胱癌患者测序数据显示其中44个基因在膀胱癌中具有预后价值（P<0.05），研究进一步基于其中18个基因成功构建了风险回归模型［曲线下面积（AUC）=0.819］，结合基因在癌与癌旁中的表达水平筛选获得MMP9、MAOA、PSMB5、HCAR及脂肪酸合成酶（FASN）作为丹参治疗膀胱癌的分子靶点，其中的FASN风险系数最高（core=0.007 412）。细胞实验表明FASN在膀胱癌细胞系J82中表达明显增高，而经过丹参（IC50=10 mg/L）处理后，膀胱癌细胞的增殖、侵袭及迁移能力均明显减弱，且FASN蛋白也表达减少。结论　本研究揭示了丹参的有效活性成分及其治疗膀胱癌的分子靶点，为临床应用丹参治疗膀胱癌提供了必要的实验依据与理论基础。

关键词:

膀胱癌, 丹参, FASN, 网络药理学

Gui Zhiming, Huang Jian, Li Shihao, Wu Haokai, Wang Qianliang.

Molecular mechanism of Salvia miltiorrhiza inhibiting FASN expression in the treatment of bladder cancer based on network pharmacology analysis [J]. International Medicine and Health Guidance News, 2023, 29(23): 3422-3429.

桂志明黄健李世豪吴昊开汪前亮.

基于网络药理学分析丹参抑制FASN表达治疗膀胱癌的分子机制研究 [J]. 国际医药卫生导报, 2023, 29(23): 3422-3429.

References

[1] Ferlay J, Soerjomataram I, Dikshit R, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012[J]. Int J Cancer,2015,136(5):E359-E386.DOI:10.1002/ijc.29210.
[2] Antoni S, Ferlay J, Soerjomataram I, et al. Bladder cancer incidence and mortality: a global overview and recent trends[J].Eur Urol,2017,71(1):96-108.DOI:10.1016/j.eururo.2016.06.010.
[3] Lobo N, Mount C, Omar K, et al. Landmarks in the treatment of muscle-invasive bladder cancer[J].Nat Rev Urol,2017,14(9):565-574.DOI:10.1038/nrurol.2017.82.
[4] Cathomas R, Lorch A, Bruins HM, et al. The 2021 updated european association of urology guidelines on metastatic urothelial carcinoma[J].Eur Urol,2022,81(1):95-103.DOI:10.1016/j.eururo.2021.09.026.
[5] 刘丽宁. 传统中药临床用药优势之一多靶点探讨[J]. 时珍国医国药,2010,21(3):745-746.DOI:10.3969/j.issn.1008-0805.2010.03.116.
[6] 徐晋红. 关于中药保护的思考[J]. 国际医药卫生导报,2005,20(3):92-94.DOI:10.3760/cma.j.issn.1007-1245.2005. 03.040.
[7] Liu S, Wang Y, Shi M, et al. SmbHLH60 and SmMYC2 antagonistically regulate phenolic acids and anthocyanins biosynthesis in Salvia miltiorrhiza[J].J Adv Res,2022,42:205-219.DOI:10.1016/j.jare.2022.02.005.
[8] Zhang X, Zhang P, An L, et al. Miltirone induces cell death in hepatocellular carcinoma cell through GSDME-dependent pyroptosis[J].Acta Pharm Sin B,2020,10(8):1397-1413.DOI:10.1016/j.apsb.2020.06.015.
[9] Wu CY, Yang YH, Lin YS, et al. Dihydroisotanshinone I induced ferroptosis and apoptosis of lung cancer cells[J].Biomed Pharmacother,2021,139:111585.DOI:10.1016/j.biopha.2021.111585.
[10] Han H, Qian C, Zong G, et al. Systemic pharmacological verification of Salvia miltiorrhiza-Ginseng Chinese herb pair in inhibiting spontaneous breast cancer metastasis[J].Biomed Pharmacother,2022,156:113897.DOI:10.1016/j.biopha.2022.113897.
[11] Han S, Bi S, Guo T, et al. Nano co-delivery of plumbagin and dihydrotanshinone I reverses immunosuppressive TME of liver cancer[J].J Control Release,2022,348:250-263.DOI:10.1016/j.jconrel.2022.05.057.
[12] Zhang R, Zhu X, Bai H, et al. Network pharmacology databases for traditional Chinese medicine: review and assessment[J].Front Pharmacol,2019,10:123.DOI:10.3389/fphar.2019.00123.
[13] Jiao W, Mi S, Sang Y, et al. Integrated network pharmacology and cellular assay for the investigation of an anti-obesity effect of 6-shogaol[J].Food Chem,2022,374:131755.DOI:10.1016/j.foodchem.2021.131755.
[14] Zhou W, Zhang H, Wang X, et al. Network pharmacology to unveil the mechanism of Moluodan in the treatment of chronic atrophic gastritis[J].Phytomedicine,2022,95:153837.DOI:10.1016/j.phymed.2021.153837.
[15] 牛海帆,马宁. 基于GEO数据库及网络药理学探究古方黄芪散治疗糖尿病肾病相关作用机制[J]. 国际医药卫生导报,2021,27(15):2242-2247.DOI:10.3760/cma.j.issn.1007- 1245.2021.15.006.
[16] Zhou J, Jiang YY, Chen H, et al. Tanshinone I attenuates the malignant biological properties of ovarian cancer by inducing apoptosis and autophagy via the inactivation of PI3K/AKT/mTOR pathway[J].Cell Prolif,2020,53(2):e12739.DOI:10.1111/cpr.12739.
[17] Chen Y, Li H, Li M, et al. Salvia miltiorrhiza polysaccharide activates T Lymphocytes of cancer patients through activation of TLRs mediated -MAPK and -NF-κB signaling pathways[J].J Ethnopharmacol,2017,200:165-173.DOI:10.1016/j.jep.2017.02.029.
[18] Cao Y, Huang B, Gao C. Salvia miltiorrhiza extract dihydrotanshinone induces apoptosis and inhibits proliferation of glioma cells[J].Bosn J Basic Med Sci,2017,17(3):235-240.DOI:10.17305/bjbms.2017.1800.
[19] Park IJ, Kim MJ, Park OJ, et al. Cryptotanshinone sensitizes DU145 prostate cancer cells to Fas(APO1/CD95)-mediated apoptosis through Bcl-2 and MAPK regulation[J].Cancer Lett,2010,298(1):88-98.DOI:10.1016/j.canlet.2010.06.006.
[20] Chen X, Zhang X, Jiang Y, et al. YAP1 activation promotes epithelial-mesenchymal transition and cell survival of renal cell carcinoma cells under shear stress[J].Carcinogenesis,2022,43(4):301-310.DOI:10.1093/carcin/bgac014.
[21] Huang Y, Yu SH, Zhen WX, et al. Tanshinone I, a new EZH2 inhibitor restricts normal and malignant hematopoiesis through upregulation of MMP9 and ABCG2[J].Theranostics,2021,11(14):6891-6904.DOI:10.7150/thno.53170.
[22] Li S, Yang K, Cao W, et al. Tanshinone IIA enhances the therapeutic efficacy of mesenchymal stem cells derived exosomes in myocardial ischemia/reperfusion injury via up-regulating miR-223-5p[J].J Control Release,2023,358:13-26.DOI:10.1016/j.jconrel.2023.04.014.
[23] Wells WA, Schwartz GN, Morganelli PM, et al. Expression of "Spot 14" (THRSP) predicts disease free survival in invasive breast cancer: immunohistochemical analysis of a new molecular marker[J].Breast Cancer Res Treat,2006,98(2):231-240.DOI:10.1007/s10549-005-9154-z.
[24] Kawamura T, Kanno R, Fujii H, et al. Expression of liver-type fatty-acid-binding protein, fatty acid synthase and vascular endothelial growth factor in human lung carcinoma[J].Pathobiology,2005,72(5):233-240.DOI:10.1159/000089417.
[25] Shah US, Dhir R, Gollin SM, et al. Fatty acid synthase gene overexpression and copy number gain in prostate adenocarcinoma[J].Hum Pathol,2006,37(4):401-409.DOI:10.1016/j.humpath.2005.11.022.
[26] Kuhajda FP. Fatty-acid synthase and human cancer: new perspectives on its role in tumor biology[J].Nutrition,2000,16(3):202-208.DOI:10.1016/s0899-9007(99)00266-x.
[27] Tian WX. Inhibition of fatty acid synthase by polyphenols[J].Curr Med Chem,2006,13(8):967-977.DOI:10.2174/092986706776361012.
[28] Kusunoki J, Kanatani A, Moller DE. Modulation of fatty acid metabolism as a potential approach to the treatment of obesity and the metabolic syndrome[J].Endocrine,2006,29(1):91-100.DOI:10.1385/ENDO:29:1:91.
[29] Xiong Q, Feng D, Wang Z, et al. Fatty acid synthase is the key regulator of fatty acid metabolism and is related to immunotherapy in bladder cancer[J].Front Immunol,2022,13:836939.DOI:10.3389/fimmu.2022.836939.
[30] Deng J, Peng M, Zhou S, et al. Metformin targets Clusterin to control lipogenesis and inhibit the growth of bladder cancer cells through SREBP-1c/FASN axis[J].Signal Transduct Target Ther,2021,6(1):98.DOI:10.1038/s41392-021-00493-8.