The Tanshinones of Salvia miltiorrhiza Bunge Suppress GES-1 Apoptosis via Inhibiting EGFR-STAT3 Signaling Pathway in vitro

Yujia Weng; Hanlei Huang; Liping Han; Zhaohuan Lou

doi:10.62767/jecacm602.6554

Authors

Yujia Weng School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, 310053 Hangzhou, Zhejiang, China; Center for Drug Safety Evaluation and Research, College of Pharmaceutical Sciences, Zhejiang University, 310053 Hangzhou, Zhejiang, China Author
Hanlei Huang School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, 310053 Hangzhou, Zhejiang, China Author
Liping Han School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, 310053 Hangzhou, Zhejiang, China Author
Zhaohuan Lou School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, 310053 Hangzhou, Zhejiang, China; Zhejiang Provincial Key Laboratory of Pharmacology Research on TCM Treatment of Hypertension and related diseases, 310053 Hangzhou, Zhejiang, China Author

DOI:

https://doi.org/10.62767/jecacm602.6554

Keywords:

Tanshinone, EGFR-STAT3 signaling pathway, Chronic atrophic gastritis, Network pharmacology

Abstract

Aim: Chronic atrophic gastritis (CAG), a precancerous lesion of gastric cancer. Current therapies face challenges including drug resistance and incomplete efficacy. Tanshinones exhibit anti-inflammatory and anti-tumor properties, yet their role in CAG remains unexplored. The present study aimed to explore the therapeutic effect and underlying mechanism of tanshinones on N-methyl-N’-nitro-N-nitrosoguanidine (MNNG)-induced CAG in vitro. Methods: The active tanshinones were screened from CNKI, PubMed and TCMSP databases. The CAG-related targets were collected from GeneCards database. Cytoscape 3.9.0 software was used to construct a compound-target network. A protein-protein interaction (PPI) network was constructed using a string database. The potential mechanism was analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Molecular docking was completed by Autodock software. An in vitro CAG model was established using the human gastric epithelial cell line GES-1, induced by MNNG. Cell viability of GES-1 was measured by Cell Counting Kit-8 (CCK-8) assay. TUNEL assay was used to evaluate cell apoptosis and protein expression of Epidermal growth factor receptor (EGFR), Signal transducer and activator of transcription 3 (STAT3) etc was determined by Western blot. Results: The compound-target network analysis found 12 active tanshinones and 42 common targets. The key targets were Cysteinyl Aspartate-specific Proteinase-3 (Caspase-3 (CASP3)), EGFR, STAT3, PTGS2, MMP2, ERBB2, and PPARG. Through enrichment analysis, 656 GO terms were enriched, whereas the top 15 pathways, including proteoglycans in cancer and apoptosis, were identified. Through molecular docking, cryptotanshinone, tanshinone IIA, dihydrotanshinone I, and tanshinone I were found to be stably bound to EGFR and STAT3. In vitro experiments indicated that tanshinones could significantly inhibit MNNG-induced GES-1 cell apoptosis, and markedly lower the protein expression of EGFR, STAT3, and Caspase-3. Conclusion: Tanshinones combination exhibited a positive effect on anti-CAG induced by MNNG in vitro, and this function is correlated with the suppression of EGFR-STAT3 signaling pathway and cell apoptosis.

References

Rodriguez-Castro KI, Franceschi M, Noto A, et al. Clinical manifestations of chronic atrophic gastritis. Acta bio-medica: Atenei Parmensis 2018; 89(8-S): 88-92.

Annibale B, Esposito G, Lahner E. A current clinical overview of atrophic gastritis. Expert Review of Gastroenterology & Hepatology 2020; 14(2): 93-102.

Correa P. Helicobacter pylori and gastric carcinogenesis. American Journal of Surgical Pathology 1995; 19 Suppl 1: S37-43.

Lahner E, Carabotti M, Annibale B. Treatment of Helicobacter pylori infection in atrophic gastritis. World Journal of Gastroenterology 2018; 24(22): 2373-2380.

Fang WJ, Zhang XY, Yang B, et al. Chinese Herbal Decoction as a Complementary Therapy for Atrophic Gastritis: A Systematic Review and Meta-Analysis. African Journal of Traditional, Complementary, and Alternative Medicines: AJTCAM 2017; 14(4): 297-319.

Su CY, Ming QL, Rahman K, et al. Salvia miltiorrhiza: Traditional medicinal uses, chemistry, and pharmacology. Chinese Journal of Natural Medicines 2015; 13(3): 163-182.

Wang X, Yang Y, Liu X, et al. Pharmacological properties of tanshinones, the natural products from Salvia miltiorrhiza. Advances in Pharmacology (San Diego, Calif.) 2020; 87: 43-70.

Xia M, Wu Y, Zhu H, Duan W. Tanshinone I induces ferroptosis in gastric cancer cells via the KDM4D/p53 pathway. Human & Experimental Toxicology 2023; 42: 9603271231216963.

Luo N, Zhang K, Li X, et al. Tanshinone IIA destabilizes SLC7A11 by regulating PIAS4-mediated SUMOylation of SLC7A11 through KDM1A, and promotes ferroptosis in breast cancer. Journal of Advanced Research 2025; 69: 313-327.

Qian J, Cao Y, Zhang J, et al. Tanshinone IIA induces autophagy in colon cancer cells through MEK/ERK/mTOR pathway. Translational Cancer Research 2020; 9(11): 6919-6928.

Lou ZH, Xia RM, Li XJ, et al. Anti-lung cancer mechanisms of diterpenoid tanshinone via endoplasmic reticulum stress-mediated apoptosis signal pathway. China Journal of Chinese Materia Medica 2018; 43(24): 4900-4907.

Cui S, Chen S, Wu Q, et al. A network pharmacology approach to investigate the anti-inflammatory mechanism of effective ingredients from Salvia miltiorrhiza. International Immunopharmacology 2020; 81: 106040.

Liu X, He H, Huang T, et al. Tanshinone IIA Protects against Dextran Sulfate Sodium- (DSS-) Induced Colitis in Mice by Modulation of Neutrophil Infiltration and Activation. Oxidative Medicine and Cellular Longevity 2016; 2016: 7916763.

Yu JR, Liu YY, Gao YY, et al. Diterpenoid tanshinones inhibit gastric cancer angiogenesis through the PI3K/Akt/mTOR signaling pathway. Journal of Ethnopharmacology 2024; 324: 117791.

Guan Z, Chen J, Li X, et al. Tanshinone IIA induces ferroptosis in gastric cancer cells through p53-mediated SLC7A11 down-regulation. Bioscience Reports 2020; 40(8): BSR20201807.

Gao H, Huang L, Ding F, et al. Simultaneous purification of dihydrotanshinone, tanshinone I, cryptotanshinone, and tanshinone IIA from Salvia miltiorrhiza and their anti-inflammatory activities investigation. Scientific Reports 2018; 8(1): 8460.

Liu L, Gao H, Wen T, et al. Tanshinone IIA attenuates AOM/DSS-induced colorectal tumorigenesis in mice via inhibition of intestinal inflammation. Pharmaceutical Biology 2021; 59(1): 89-96.

Shen L, Lou Z, Zhang G, et al. Diterpenoid Tanshinones, the extract from Danshen (Radix Salviae Miltiorrhizae) induced apoptosis in nine human cancer cell lines. Journal of Traditional Chinese Medicine 2016; 36(4): 514-521.

Lou ZH, Xia RM, Li XJ, et al. Anti-lung cancer mechanisms of diterpenoid tanshinone via endoplasmic reticulum stress-mediated apoptosis signal pathway. Zhongguo Zhong Yao Za Zhi 2018; 43(24): 4900-4907.

Zhang R, Zhu X, Bai H, et al. Network Pharmacology Databases for Traditional Chinese Medicine: Review and Assessment. Frontiers in Pharmacology 2019; 10: 123.

Li S, Zhang B. Traditional Chinese medicine network pharmacology: theory, methodology and application. Chinese Journal of Natural Medicines 2013; 11(2): 110-120.

Guo B, Zhao C, Zhang C, et al. Elucidation of the anti-inflammatory mechanism of Er Miao San by integrative approach of network pharmacology and experimental verification. Pharmacological Research 2022; 175: 106000.

Guo H, Zeng H, Fu C, et al. Identification of Sitogluside as a Potential Skin-Pigmentation-Reducing Agent through Network Pharmacology. Oxidative Medicine and Cellular Longevity 2021; 2021: 4883398.

Wen JX, Tong YL, Ma X, et al. Therapeutic effects and potential mechanism of dehydroevodiamine on N-methyl-N'-nitro-N-nitrosoguanidine-induced chronic atrophic gastritis. Phytomedicine 2021; 91: 153619.

Ru J, Li P, Wang J, et al. TCMSP: a database of systems pharmacology for drug discovery from herbal medicines. Journal of Cheminformatics 2014; 6: 13.

Zhou Y, Zhou B, Pache L, et al. Metascape provides a biologist-oriented resource for the analysis of systems-level datasets. Nature Communications 2019; 10(1): 1523.

Aoyagi K, Kohfuji K, Yano S, et al. Morphological change in the MNNG-treated rat gastric mucosa. The Kurume Medical Journal 2000; 47(1): 31-36.

Lahner E, Esposito G, Pilozzi E, et al. Occurrence of gastric cancer and carcinoids in atrophic gastritis during prospective long-term follow up. Scandinavian Journal of Gastroenterology 2015; 50(7): 856-865.

Asano T. Drug Resistance in Cancer Therapy and the Role of Epigenetics. Journal of Nippon Medical School 2020; 87(5): 244-251.

Ma S, Li Y, Guo R, et al. The Efficacy of Morodan in Combination with Rabeprazole for the Treatment of Chronic Gastritis and its Impact on Gastric Mucosal Repair. Alternative Therapies in Health and Medicine 2023; 29(6): 306-310.

Shinkai M, Imano M, Hiraki Y, et al. Combination Chemotherapy Including Intraperitoneal (IP) Administration of Paclitaxel (PTX) followed by PTX, CDDP and S-1 Triplet Chemotherapy for CY1P0 Gastric Cancer. Gan to kagaku Ryoho. Cancer & Chemotherapy 2017; 44(12): 1355-1357.

Li CJ, Chu CY, Huang LH, et al. Synergistic anticancer activity of triptolide combined with cisplatin enhances apoptosis in gastric cancer in vitro and in vivo. Cancer Letters 2012; 319(2): 203-213.

He X, Zhang T, Wu L, et al. The effect of trifluridine/tipiracil for patients with heavily pretreated metastatic gastric cancer: A protocol for systematic review and meta-analysis. Medicine (Baltimore) 2021; 100(2): e24110.

Zhou Z, Chen B, Chen S, et al. Applications of Network Pharmacology in Traditional Chinese Medicine Research. Evidence-Based Complementary and Alternative Medicine: eCAM 2020; 2020: 1646905.

Quesnelle KM, Boehm AL, Grandis JR. STAT-mediated EGFR signaling in cancer. Journal of Cellular Biochemistry 2007; 102(2): 311-319.

Wu M, Zhang P. EGFR-mediated autophagy in tumourigenesis and therapeutic resistance. Cancer Letter 2020; 469: 207-216.

Li SY, Hou LZ, Gao YX, et al. FIP-nha, a fungal immunomodulatory protein from Nectria haematococca, induces apoptosis and autophagy in human gastric cancer cells via blocking the EGFR-mediated STAT3/Akt signaling pathway. Food Chemistry 2022; 4: 100091.

Wee P, Wang Z. Epidermal Growth Factor Receptor Cell Proliferation Signaling Pathways. Cancers (Basel) 2017; 9(5): 52.

Rajaram P, Chandra P, Ticku S, et al. Epidermal growth factor receptor: Role in human cancer. Indian Journal of Dental Research: Official Publication of Indian Society for Dental Research 2017; 28(6): 687-694.

Lin HY, Zhao Y, Yu JN, et al. Effects of traditional Chinese medicine Wei-Wei-Kang-Granule on the expression of EGFR and NF-KB in chronic atrophic gastritis rats. African Journal of Traditional, Complementary, and Alternative Medicines: AJTCAM 2012; 9(1): 1-7.

Chen X, Wang R, Bao C, et al. Palmatine ameliorates Helicobacter pylori-induced chronic atrophic gastritis by inhibiting MMP-10 through ADAM17/EGFR. European Journal of Pharmacology 2020; 882: 173267.

Darnell JE, Jr. STATs and gene regulation. Science 1997; 277(5332): 1630-1635.

Loh CY, Arya A, Naema AF, et al. Signal Transducer and Activator of Transcription (STATs) Proteins in Cancer and Inflammation: Functions and Therapeutic Implication. Frontiers in Oncology 2019; 9: 48.

Ishii Y, Shibata W, Sugimori M, et al. Activation of Signal Transduction and Activator of Transcription 3 Signaling Contributes to Helicobacter-Associated Gastric Epithelial Proliferation and Inflammation. Gastroenterology Research and Practice 2018; 2018: 9050715.

Hu Y, Zeng T, Xiao Z, et al. Immunological role and underlying mechanisms of B7-H6 in tumorigenesis. Clinica Chimica Acta; International Journal of Clinical Chemistry 2020; 502: 191-198.

Goldar S, Khaniani MS, Derakhshan SM, et al. Molecular mechanisms of apoptosis and roles in cancer development and treatment. Asian Pacific Journal of Cancer Prevention: APJCP 2015; 16(6): 2129-2144.

Xia HH, Talley NJ. Apoptosis in gastric epithelium induced by Helicobacter pylori infection: implications in gastric carcinogenesis. American Journal of Gastroenterology 2001; 96(1): 16-26.