Transforming Growth Factor Beta2 Promotes Migration and Inhibits the Proliferation of Gastric Cancer Cells by Regulating the pSmad2/3-NDRG1 Signaling Pathway

Abstract

Transforming growth factor beta2 (TGFβ2) is upregulated in gastric cancer (GC), playing a crucial role in driving its progression. However, the biological effects of TGFβ2 in GC metastasis and proliferation remain not fully understood. Our study reveals that TGFβ2 enhances N-myc downstream-regulated gene 1 (NDRG1) protein expression by activating the TGFβR/Smad2/3-dependent pathway, accelerating GC progression. TGFβ2 knockdown downregulates NDRG1 by inhibiting the TGFβR/Smad2/3 signaling pathway, which in turn inhibits GC cell migration and epithelial–mesenchymal transition (EMT) but stimulates proliferation. Both TGFβ2 upregulation and NDRG1 upregulation enhance GC cell migration in vitro and promote lung metastasis in mouse models. Interfering with NDRG1 reverses TGFβ2-induced migration, and inhibiting Smad2/3 or TGFβR reverses TGFβ2-induced NDRG1 upregulation and GC cell migration. Clinical sample analysis shows high TGFβ2 and NDRG1 expression in GC, associated with poor prognosis. Our study reveals that TGFβ2 upregulates NDRG1 via the TGFβR/Smad2/3 pathway, driving GC progression and highlighting the potential role of the TGFβ2NDRG1 axis in GC-targeted therapies.

Keywords

gastric cancer / metastasis / N-myc downstream-regulated gene 1 / proliferation / transforming growth factor beta2

References

[1] H. Sung, J. Ferlay, R. L. Siegel, et al., “Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries,” CA: A Cancer Journal for Clinicians 71, no. 3 (2021): 209-249.

[2] Chinese Gastric Cancer Association of Chinese Anti-Cancer Association, Chinese Society of Upper Gastrointestinal Surgeons of Chinese Medical Doctor Association, and Chinese Health Risk Management Collaboration-Gastric Cancer Group, “[Chinese Guideline on Risk Management of Gastric Cancer in the General Public(2023 Edition)],” Zhonghua Yi Xue Za Zhi 103, no. 36 (2023): 2837-2849.

[3] C. Li, Y. Zheng, Z. Shi, et al. “Perioperative Camrelizumab (C) Combined With Rivoceranib (R) and Chemotherapy (Chemo) Versus Chemo for Locally Advanced Resectable Gastric or Gastroesophageal Junction (G/GEJ) Adenocarcinoma: The First Interim Analysis of a Randomized, Phase III Trial (DRAGON IV),” Annals of Oncology 34, no. supplement S2 (2023): S852-S886.

[4] D. Li, X. Guo, K. Yang, et al., “EpCAM-Targeting CAR-T Cell Immunotherapy Is Safe and Efficacious for Epithelial Tumors,” Science Advances 9, no. 48 (2023): eadg9721.

[5] C. Allemani, T. Matsuda, V. Di Carlo, et al., “Global Surveillance of Trends in Cancer Survival 2000-14 (CONCORD-3): Analysis of Individual Records for 37 513 025 Patients Diagnosed With One of 18 Cancers From 322 Population-Based Registries in 71 Countries,” Lancet 391, no. 10125 (2018): 1023-1075.

[6] Y. Xia, Z. Wei, W. Huang, X. Wei, and Y. He, “Trim47 Overexpression Correlates With Poor Prognosis in Gastric Cancer,” Neoplasma 68, no. 2 (2021): 307-316.

[7] A. W. Lambert, D. R. Pattabiraman, and R. A. Weinberg, “Emerging Biological Principles of Metastasis,” Cell 168, no. 4 (2017): 670-691.

[8] S. Gerstberger, Q. Jiang, and K. Ganesh, “Metastasis,” Cell 186, no. 8 (2023): 1564-1579.

[9] J. Massagué and K. Ganesh, “Metastasis-Initiating Cells and Ecosystems,” Cancer Discovery 11, no. 4 (2021): 971-994.

[10] D. F. Quail and J. A. Joyce, “Microenvironmental Regulation of Tumor Progression and Metastasis,” Nature Medicine 19, no. 11 (2013): 1423-1437.

[11] D. Kimelman and M. Kirschner, “Synergistic Induction of Mesoderm by FGF and TGF-Beta and the Identification of an mRNA Coding for FGF in the Early Xenopus Embryo,” Cell 51, no. 5 (1987): 869-877.

[12] D. Sun, C. R. Vanderburg, G. S. Odierna, and E. D. Hay, “TGFbeta3 Promotes Transformation of Chicken Palate Medial Edge Epithelium to Mesenchyme In Vitro,” Development 125, no. 1 (1998): 95-105.

[13] T. Sun, Z. Huang, W. C. Liang, et al., “TGFβ2 and TGFβ3 Isoforms Drive Fibrotic Disease Pathogenesis,” Science Translational Medicine 13, no. 605 (2021): eabe0407.

[14] R. Derynck and Y. E. Zhang, “Smad-Dependent and Smad-Independent Pathways in TGF-Beta family Signalling,” Nature 425, no. 6958 (2003): 577-584.

[15] M. E. Engel, M. A. McDonnell, B. K. Law, and H. L. Moses, “Interdependent SMAD and JNK Signaling in Transforming Growth Factor-Beta-Mediated Transcription,” Journal of Biological Chemistry 274, no. 52 (1999): 37413-37420.

[16] L. Yu, M. C. Hébert, and Y. E. Zhang, “TGF-Beta Receptor-Activated p38 MAP Kinase Mediates Smad-Independent TGF-Beta Responses,” EMBO Journal 21, no. 14 (2002): 3749-3759.

[17] S. Edlund, M. Landström, C. H. Heldin, and P. Aspenström, “Transforming Growth Factor-Beta-Induced Mobilization of Actin Cytoskeleton Requires Signaling by Small GTPases Cdc42 and RhoA,” Molecular Biology of the Cell 13, no. 3 (2002): 902-914.

[18] A. L. Bishop and A. Hall, “Rho GTPases and Their Effector Proteins,” Biochemical Journal 348, no. 2 (2000): 241-255.

[19] L. Luo, N. Xu, W. Fan, et al., “The TGFβ2-Snail1-miRNATGFβ2 Circuitry Is Critical for the Development of Aggressive Functions in Breast Cancer,” Clinical and Translational Medicine 14, no. 2 (2024): e1558.

[20] J. Massagué, “TGFbeta in Cancer,” Cell 134, no. 2 (2008): 215-230.

[21] N. Dünker, K. Schmitt, and K. Krieglstein, “TGF-Beta Is Required for Programmed Cell Death in Interdigital Webs of the Developing Mouse Limb,” Mechanisms of Development 113, no. 2 (2002): 111-120.

[22] J. J. Letterio and A. B. Roberts, “Regulation of Immune Responses by TGF-Beta,” Annual Review of Immunology 16 (1998): 137-161.

[23] Z. Yang, X. Wu, J. Shen, et al., “Ginsenoside Rh1 Regulates Gastric Cancer Cell Biological Behaviours and Transplanted Tumour Growth in Nude Mice via the TGF-β/Smad Pathway,” Clinical and Experimental Pharmacology & Physiology 49, no. 12 (2022): 1270-1280.

[24] Z. Wang, Y. Yang, Y. Cui, et al., “Tumor-Associated Macrophages Regulate Gastric Cancer Cell Invasion and Metastasis Through TGFβ2/NF-κB/Kindlin-2 Axis,” Journal of Cancer Research 34, no. 1 (2022): 66.

[25] J. Massagué, S. W. Blain, and R. S. Lo, “TGFbeta Signaling in Growth Control, Cancer, and Heritable Disorders,” Cell 103, no. 2 (2000): 295-309.

[26] J. Li and L. Kretzner, “The Growth-Inhibitory Ndrg1 Gene Is a Myc Negative Target in Human Neuroblastomas and Other Cell Types With Overexpressed N- or c-myc,” Molecular and Cellular Biochemistry 250, no. 1-2 (2003): 91-105.

[27] W. Cui, D. J. Fowlis, S. Bryson, et al., “TGFbeta1 Inhibits the Formation of Benign Skin Tumors, but Enhances Progression to Invasive Spindle Carcinomas in Transgenic Mice,” Cell 86, no. 4 (1996): 531-542.

[28] J. J. Yin, K. Selander, J. M. Chirgwin, et al., “TGF-Beta Signaling Blockade Inhibits PTHrP Secretion by Breast Cancer Cells and Bone Metastases Development,” Journal of Clinical Investigation 103, no. 2 (1999): 197-206.

[29] K. Polyak, “Negative Regulation of Cell Growth by TGF Beta,” Biochimica et Biophysica Acta 1242, no. 3 (1996): 185-199.

[30] K. Takaku, M. Oshima, H. Miyoshi, M. Matsui, M. F. Seldin, and M. M. Taketo, “Intestinal Tumorigenesis in Compound Mutant Mice of Both Dpc4 (Smad4) and Apc Genes,” Cell 92, no. 5 (1998): 645-656.

[31] Y. Shi, Y. F. Wang, L. Jayaraman, H. Yang, J. Massagué, and N. P. Pavletich, “Crystal Structure of a Smad MH1 Domain Bound to DNA: Insights on DNA Binding in TGF-Beta Signaling,” Cell 94, no. 5 (1998): 585-594.

[32] M. Stankic, S. Pavlovic, Y. Chin, et al., “TGF-β-Id1 Signaling Opposes Twist1 and Promotes Metastatic Colonization via a Mesenchymal-to-Epithelial Transition,” Cell Reports 5, no. 5 (2013): 1228-1242.

[33] J. P. Thiery, H. Acloque, R. Y. Huang, and M. A. Nieto, “Epithelial-Mesenchymal Transitions in Development and Disease,” Cell 139, no. 5 (2009): 871-890.

[34] Y. Kimura, H. Ohzawa, H. Miyato, et al., “Intraperitoneal Transfer of microRNA-29b-Containing Small Extracellular Vesicles Can Suppress Peritoneal Metastases of Gastric Cancer,” Cancer Science 114, no. 7 (2023): 2939-2950.

[35] Z. Chen, D. Zhang, F. Yue, M. Zheng, Z. Kovacevic, and D. R. Richardson, “The Iron Chelators Dp44mT and DFO Inhibit TGF-β-induced Epithelial-Mesenchymal Transition via Up-Regulation of N-Myc Downstream-Regulated Gene 1 (NDRG1),” Journal of Biological Chemistry 287, no. 21 (2012): 17016-17028.

[36] Y. Inagaki, W. Tang, H. L. Xu, et al., “Localization of N-myc Downstream-Regulated Gene 1 in Gastric Cancer Tissue,” Digestive and Liver Disease 41, no. 2 (2009): 96-103.

[37] R. J. Guan, H. L. Ford, Y. Fu, Y. Li, L. M. Shaw, and A. B. Pardee, “Drg-1 as a Differentiation-Related, Putative Metastatic Suppressor Gene in Human Colon Cancer,” Cancer Research 60, no. 3 (2000): 749-755.

[38] N. van Belzen, W. N. Dinjens, M. P. Diesveld, et al., “A Novel Gene Which Is Up-Regulated During Colon Epithelial Cell Differentiation and Down-Regulated in Colorectal Neoplasms,” Laboratory Investigation 77, no. 1 (1997): 85-92.

[39] M. S. Chua, H. Sun, S. T. Cheung, et al., “Overexpression of NDRG1 Is an Indicator of Poor Prognosis in Hepatocellular Carcinoma,” Modern Pathology 20, no. 1 (2007): 76-83.

[40] B. Strzelczyk, A. Szulc, R. Rzepko, et al., “Identification of High-Risk Stage II Colorectal Tumors by Combined Analysis of the NDRG1 Gene Expression and the Depth of Tumor Invasion,” Annals of Surgical Oncology 16, no. 5 (2009): 1287-1294.

[41] Y. Maruyama, M. Ono, A. Kawahara, et al., “Tumor Growth Suppression in Pancreatic Cancer by a Putative Metastasis Suppressor Gene Cap43/NDRG1/Drg-1 Through Modulation of Angiogenesis,” Cancer Research 66, no. 12 (2006): 6233-6242.

[42] R. Gerhard, S. Nonogaki, J. H. Fregnani, F. A. Soares, and M. A. Nagai, “NDRG1 Protein Overexpression in Malignant Thyroid Neoplasms,” Clinics 65, no. 8 (2010): 757-762.

[43] S. Nishio, K. Ushijima, N. Tsuda, et al., “Cap43/NDRG1/Drg-1 Is a Molecular Target for Angiogenesis and a Prognostic Indicator in Cervical Adenocarcinoma,” Cancer Letters 264, no. 1 (2008): 36-43.

[44] J. Y. Shin, J. D. Beckett, R. Bagirzadeh, et al., “Epigenetic Activation and Memory at a TGFB2 Enhancer in Systemic Sclerosis,” Science Translational Medicine 11, no. 497 (2019): eaaw0790.

[45] J. L. Gulley, J. Schlom, M. H. Barcellos-Hoff, et al., “Dual Inhibition of TGF-β and PD-L1: A Novel Approach to Cancer Treatment,” Molecular Oncology 16, no. 11 (2022): 2117-2134.

[46] J. P. Annes, J. S. Munger, and D. B. Rifkin, “Making Sense of Latent TGFbeta Activation,” Journal of Cell Science 116, no. 2 (2003): 217-224.

RIGHTS & PERMISSIONS

2025 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.