Skip to main content

Advertisement

SpringerLink
Log in

Secretome analysis reveals reduced expression of COL4A2 in hypoxic cancer-associated fibroblasts with a tumor-promoting function in gastric cancer

  • Research
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Background

Cancer-associated fibroblasts (CAFs) are major components of the tumor microenvironment (TME). Hypoxic TME is known to promote tumor progression. However, how a hypoxic condition regulates CAFs remains elusive.

Methods

To investigate the underlying mechanism involved in the regulation of gastric cancer (GC) progression by hypoxic CAFs, we performed secretome profiling. Normoxic or hypoxic CAFs conditioned media (CM) were filter-concentrated and in-gel trypsin digested. Resulting peptides were analyzed with LC–MS/MS.

Results

We observed that CM derived from hypoxic CAFs could promote migration of a panel of GC cell lines (AGS, SNU668, SNU638). Mass spectrometry analysis of hypoxic or normoxic CAFs CM identified 1595 proteins, of which 19 proteins (10 upregulated and 9 downregulated) were differentially expressed in the hypoxic secretome. We focused on COL4A2, whose expression was significantly decreased in hypoxic CAFs in HIF-1α-independent manner. Silencing of COL4A2 expression in normoxic CAFs phenocopied the effect of hypoxic CAFs in promoting GC cell migration.

Conclusions

The reduced expression of COL4A2 in a hypoxic environment might be associated with the tumor-promoting role of hypoxic CAFs in GC.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Al Tameemi W, Dale TP, Al-Jumaily RMK, Forsyth NR (2019) Hypoxia-modified cancer cell metabolism. Front Cell Dev Biol 7:4

    PubMed  PubMed Central  Google Scholar 

  • Bai Y, Kim JY, Watters JM, Fang B, Kinose F, Song L, Koomen JM, Teer JK, Fisher K, Chen YA, Rix U, Haura EB (2014) Adaptive responses to dasatinib-treated lung squamous cell cancer cells harboring DDR2 mutations. Cancer Res 74(24):7217–7228

    CAS  PubMed  PubMed Central  Google Scholar 

  • Benjamini Y, Heller R, Yekutieli D (2009) Selective inference in complex research. Philos Trans A Math Phys Eng Sci 367(1906):4255–4271

    PubMed  PubMed Central  Google Scholar 

  • Chang YS, Kong G, Sun S, Liu D, El-Naggar AK, Khuri FR, Hong WK, Lee HY (2002) Clinical significance of insulin-like growth factor-binding protein-3 expression in stage I non-small cell lung cancer. Clin Cancer Res 8(12):3796–3802

    CAS  PubMed  Google Scholar 

  • Chen WT, Huang CJ, Wu MT, Yang SF, Su YC, Chai CY (2005) Hypoxia-inducible factor-1alpha is associated with risk of aggressive behavior and tumor angiogenesis in gastrointestinal stromal tumor. Jpn J Clin Oncol 35(4):207–213

    PubMed  Google Scholar 

  • Chen YQ, Zhao CL, Li W (2009) Effect of hypoxia-inducible factor-1alpha on transcription of survivin in non-small cell lung cancer. J Exp Clin Cancer Res 28:29

    PubMed  PubMed Central  Google Scholar 

  • Choi SB, Park JB, Song TJ, Choi SY (2011) Molecular mechanism of HIF-1-independent VEGF expression in a hepatocellular carcinoma cell line. Int J Mol Med 28(3):449–454

    CAS  PubMed  Google Scholar 

  • Cirri P, Chiarugi P (2012) Cancer-associated-fibroblasts and tumour cells: a diabolic liaison driving cancer progression. Cancer Metastasis Rev 31(1–2):195–208

    PubMed  Google Scholar 

  • Distler JH, Jungel A, Pileckyte M, Zwerina J, Michel BA, Gay RE, Kowal-Bielecka O, Matucci-Cerinic M, Schett G, Marti HH, Gay S, Distler O (2007) Hypoxia-induced increase in the production of extracellular matrix proteins in systemic sclerosis. Arthritis Rheum 56(12):4203–4215

    CAS  PubMed  Google Scholar 

  • Dong Z, Venkatachalam MA, Wang J, Patel Y, Saikumar P, Semenza GL, Force T, Nishiyama J (2001) Up-regulation of apoptosis inhibitory protein IAP-2 by hypoxia. Hif-1-independent mechanisms. J Biol Chem 276(22):18702–18709

    CAS  PubMed  Google Scholar 

  • Feng Z, Qiao R, Ren Z, Hou X, Feng J, He X, Chen D (2020) Could CTSK and COL4A2 be specific biomarkers of poor prognosis for patients with gastric cancer in Asia?-a microarray analysis based on regional population. J Gastrointest Oncol 11(2):386–401

    PubMed  PubMed Central  Google Scholar 

  • Finger EC, Giaccia AJ (2010) Hypoxia, inflammation, and the tumor microenvironment in metastatic disease. Cancer Metastasis Rev 29(2):285–293

    CAS  PubMed  PubMed Central  Google Scholar 

  • Gilkes DM, Xiang L, Lee SJ, Chaturvedi P, Hubbi ME, Wirtz D, Semenza GL (2014) Hypoxia-inducible factors mediate coordinated RhoA-ROCK1 expression and signaling in breast cancer cells. Proc Natl Acad Sci U S A 111(3):E384-393

    CAS  PubMed  Google Scholar 

  • Hintz RL, Bock S, Thorsson AV, Bovens J, Powell DR, Jakse G, Petrides PE (1991) Expression of the insulin like growth factor-binding protein 3 (IGFBP-3) gene is increased in human renal carcinomas. J Urol 146(4):1160–1163

    CAS  PubMed  Google Scholar 

  • Huijbers IJ, Iravani M, Popov S, Robertson D, Al-Sarraj S, Jones C, Isacke CM (2010) A role for fibrillar collagen deposition and the collagen internalization receptor endo180 in glioma invasion. PLoS ONE 5(3):e9808

    PubMed  PubMed Central  Google Scholar 

  • Hwang-Bo J, Yoo KH, Park JH, Jeong HS, Chung IS (2012) Recombinant canstatin inhibits angiopoietin-1-induced angiogenesis and lymphangiogenesis. Int J Cancer 131(2):298–309

    PubMed  Google Scholar 

  • Kalluri R (2003) Basement membranes: structure, assembly and role in tumour angiogenesis. Nat Rev Cancer 3(6):422–433

    CAS  PubMed  Google Scholar 

  • Kauppila S, Stenback F, Risteli J, Jukkola A, Risteli L (1998) Aberrant type I and type III collagen gene expression in human breast cancer in vivo. J Pathol 186(3):262–268

    CAS  PubMed  Google Scholar 

  • Kim JY, Lee JY (2017) Targeting tumor adaption to chronic hypoxia: implications for drug resistance, and how it can be overcome. Int J Mol Sci 18(9):1854

    PubMed  PubMed Central  Google Scholar 

  • Kim JW, Evans C, Weidemann A, Takeda N, Lee YS, Stockmann C, Branco-Price C, Brandberg F, Leone G, Ostrowski MC, Johnson RS (2012) Loss of fibroblast HIF-1alpha accelerates tumorigenesis. Cancer Res 72(13):3187–3195

    CAS  PubMed  PubMed Central  Google Scholar 

  • Koong AC, Denko NC, Hudson KM, Schindler C, Swiersz L, Koch C, Evans S, Ibrahim H, Le QT, Terris DJ, Giaccia AJ (2000) Candidate genes for the hypoxic tumor phenotype. Cancer Res 60(4):883–887

    CAS  PubMed  Google Scholar 

  • Kugeratski FG, Atkinson SJ, Neilson LJ, Lilla S, Knight JRP, Serneels J, Juin A, Ismail S, Bryant DM, Markert EK, Machesky LM, Mazzone M, Sansom OJ, Zanivan S (2019) Hypoxic cancer-associated fibroblasts increase NCBP2-AS2/HIAR to promote endothelial sprouting through enhanced VEGF signaling. Sci Signal. https://doi.org/10.1126/scisignal.aan8247

    Article  PubMed  PubMed Central  Google Scholar 

  • Lee HS, Lee HK, Kim HS, Yang HK, Kim WH (2003) Tumour suppressor gene expression correlates with gastric cancer prognosis. J Pathol 200(1):39–46

    CAS  PubMed  Google Scholar 

  • Lee D, Ham IH, Son SY, Han SU, Kim YB, Hur H (2017) Intratumor stromal proportion predicts aggressive phenotype of gastric signet ring cell carcinomas. Gastric Cancer 20(4):591–601

    CAS  PubMed  Google Scholar 

  • Lee SJ, Choi KM, Bang G, Park SG, Kim EB, Choi JW, Chung YH, Kim J, Lee SG, Kim E, Kim JY (2021) Identification of nucleolin as a novel AEG-1-interacting protein in breast cancer via interactome profiling. Cancers (basel) 13:2842

    CAS  PubMed  Google Scholar 

  • Lei J, Ma J, Ma Q, Li X, Liu H, Xu Q, Duan W, Sun Q, Xu J, Wu Z, Wu E (2013) Hedgehog signaling regulates hypoxia induced epithelial to mesenchymal transition and invasion in pancreatic cancer cells via a ligand-independent manner. Mol Cancer 12:66

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liu Q, Jiang J, Zhang X, Zhang M, Fu Y (2021) Comprehensive analysis of IGFBPs as biomarkers in gastric cancer. Front Oncol 11:723131

    CAS  PubMed  PubMed Central  Google Scholar 

  • Madsen CD, Pedersen JT, Venning FA, Singh LB, Moeendarbary E, Charras G, Cox TR, Sahai E, Erler JT (2015) Hypoxia and loss of PHD2 inactivate stromal fibroblasts to decrease tumour stiffness and metastasis. EMBO Rep 16(10):1394–1408

    CAS  PubMed  PubMed Central  Google Scholar 

  • Magnon C, Galaup A, Mullan B, Rouffiac V, Bouquet C, Bidart JM, Griscelli F, Opolon P, Perricaudet M (2005) Canstatin acts on endothelial and tumor cells via mitochondrial damage initiated through interaction with alphavbeta3 and alphavbeta5 integrins. Cancer Res 65(10):4353–4361

    CAS  PubMed  Google Scholar 

  • Mamlouk S, Kalucka J, Singh RP, Franke K, Muschter A, Langer A, Jakob C, Gassmann M, Baretton GB, Wielockx B (2014) Loss of prolyl hydroxylase-2 in myeloid cells and T-lymphocytes impairs tumor development. Int J Cancer 134(4):849–858

    CAS  PubMed  Google Scholar 

  • Mazzone M, Dettori D, de Oliveira RL, Loges S, Schmidt T, Jonckx B, Tian YM, Lanahan AA, Pollard P, de Almodovar CR, De Smet F, Vinckier S, Aragones J, Debackere K, Luttun A, Wyns S, Jordan B, Pisacane A, Gallez B, Lampugnani MG, Dejana E, Simons M, Ratcliffe P, Maxwell P, Carmeliet P (2009) Heterozygous deficiency of PHD2 restores tumor oxygenation and inhibits metastasis via endothelial normalization. Cell 136(5):839–851

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mizukami Y, Li J, Zhang X, Zimmer MA, Iliopoulos O, Chung DC (2004) Hypoxia-inducible factor-1-independent regulation of vascular endothelial growth factor by hypoxia in colon cancer. Cancer Res 64(5):1765–1772

    CAS  PubMed  Google Scholar 

  • Monboisse JC, Oudart JB, Ramont L, Brassart-Pasco S, Maquart FX (2014) Matrikines from basement membrane collagens: a new anti-cancer strategy. Biochim Biophys Acta 1840(8):2589–2598

    CAS  PubMed  Google Scholar 

  • Mundel TM, Kalluri R (2007) Type IV collagen-derived angiogenesis inhibitors. Microvasc Res 74(2–3):85–89

    CAS  PubMed  PubMed Central  Google Scholar 

  • Natsuizaka M, Naganuma S, Kagawa S, Ohashi S, Ahmadi A, Subramanian H, Chang S, Nakagawa KJ, Ji X, Liebhaber SA, Klein-Szanto AJ, Nakagawa H (2012) Hypoxia induces IGFBP3 in esophageal squamous cancer cells through HIF-1alpha-mediated mRNA transcription and continuous protein synthesis. FASEB J 26(6):2620–2630

    CAS  PubMed  PubMed Central  Google Scholar 

  • Nissen NI, Karsdal M, Willumsen N (2019) Collagens and Cancer associated fibroblasts in the reactive stroma and its relation to Cancer biology. J Exp Clin Cancer Res 38(1):115

    PubMed  PubMed Central  Google Scholar 

  • Okada M, Yamawaki H (2019) A current perspective of canstatin, a fragment of type IV collagen alpha 2 chain. J Pharmacol Sci 139(2):59–64

    CAS  PubMed  Google Scholar 

  • Panka DJ, Mier JW (2003) Canstatin inhibits Akt activation and induces Fas-dependent apoptosis in endothelial cells. J Biol Chem 278(39):37632–37636

    CAS  PubMed  Google Scholar 

  • Pankov R, Yamada KM (2002) Fibronectin at a glance. J Cell Sci 115(Pt 20):3861–3863

    CAS  PubMed  Google Scholar 

  • Peng J, Wang X, Ran L, Song J, Luo R, Wang Y (2018) Hypoxia-Inducible factor 1alpha regulates the transforming growth factor beta1/SMAD family member 3 pathway to promote breast cancer progression. J Breast Cancer 21(3):259–266

    PubMed  PubMed Central  Google Scholar 

  • Piret JP, Mottet D, Raes M, Michiels C (2002) CoCl2, a chemical inducer of hypoxia-inducible factor-1, and hypoxia reduce apoptotic cell death in hepatoma cell line HepG2. Ann N Y Acad Sci 973:443–447

    CAS  PubMed  Google Scholar 

  • Rong Y, Hu F, Huang R, Mackman N, Horowitz JM, Jensen RL, Durden DL, Van Meir EG, Brat DJ (2006) Early growth response gene-1 regulates hypoxia-induced expression of tissue factor in glioblastoma multiforme through hypoxia-inducible factor-1-independent mechanisms. Cancer Res 66(14):7067–7074

    CAS  PubMed  PubMed Central  Google Scholar 

  • Roth JM, Akalu A, Zelmanovich A, Policarpio D, Ng B, MacDonald S, Formenti S, Liebes L, Brooks PC (2005) Recombinant alpha2(IV)NC1 domain inhibits tumor cell-extracellular matrix interactions, induces cellular senescence, and inhibits tumor growth in vivo. Am J Pathol 166(3):901–911

    CAS  PubMed  PubMed Central  Google Scholar 

  • Roy R, Yang J, Moses MA (2009) Matrix metalloproteinases as novel biomarkers and potential therapeutic targets in human cancer. J Clin Oncol 27(31):5287–5297

    CAS  PubMed  PubMed Central  Google Scholar 

  • Russo AE, Strong VE (2019) Gastric cancer etiology and management in asia and the west. Annu Rev Med 70:353–367

    CAS  PubMed  Google Scholar 

  • Sahai E, Astsaturov I, Cukierman E, DeNardo DG, Egeblad M, Evans RM, Fearon D, Greten FR, Hingorani SR, Hunter T, Hynes RO, Jain RK, Janowitz T, Jorgensen C, Kimmelman AC, Kolonin MG, Maki RG, Powers RS, Pure E, Ramirez DC, Scherz-Shouval R, Sherman MH, Stewart S, Tlsty TD, Tuveson DA, Watt FM, Weaver V, Weeraratna AT, Werb Z (2020) A framework for advancing our understanding of cancer-associated fibroblasts. Nat Rev Cancer 20(3):174–186

    CAS  PubMed  PubMed Central  Google Scholar 

  • Stoeltzing O, McCarty MF, Wey JS, Fan F, Liu W, Belcheva A, Bucana CD, Semenza GL, Ellis LM (2004) Role of hypoxia-inducible factor 1alpha in gastric cancer cell growth, angiogenesis, and vessel maturation. J Natl Cancer Inst 96(12):946–956

    CAS  PubMed  Google Scholar 

  • Takahashi R, Tanaka S, Hiyama T, Ito M, Kitadai Y, Sumii M, Haruma K, Chayama K (2003) Hypoxia-inducible factor-1alpha expression and angiogenesis in gastrointestinal stromal tumor of the stomach. Oncol Rep 10(4):797–802

    CAS  PubMed  Google Scholar 

  • Takaoka M, Harada H, Andl CD, Oyama K, Naomoto Y, Dempsey KL, Klein-Szanto AJ, El-Deiry WS, Grimberg A, Nakagawa H (2004) Epidermal growth factor receptor regulates aberrant expression of insulin-like growth factor-binding protein 3. Cancer Res 64(21):7711–7723

    CAS  PubMed  PubMed Central  Google Scholar 

  • Tam SY, Wu VWC, Law HKW (2020) Hypoxia-induced epithelial-mesenchymal transition in Cancers: HIF-1alpha and beyond. Front Oncol 10:486

    PubMed  PubMed Central  Google Scholar 

  • Tanjore H, Kalluri R (2006) The role of type IV collagen and basement membranes in cancer progression and metastasis. Am J Pathol 168(3):715–717

    CAS  PubMed  PubMed Central  Google Scholar 

  • Thrift AP, El-Serag HB (2020) Burden of gastric Cancer. Clin Gastroenterol Hepatol 18(3):534–542

    PubMed  Google Scholar 

  • Wang SS, Wuputra K, Liu CJ, Lin YC, Chen YT, Chai CY, Lin CS, Kuo KK, Tsai MH, Wang SW, Chen KK, Miyoshi H, Nakamura Y, Saito S, Hanafusa T, Wu DC, Lin CS, Yokoyama KK (2016) Oncogenic function of the homeobox A13-long noncoding RNA HOTTIP-insulin growth factor-binding protein 3 axis in human gastric cancer. Oncotarget 7(24):36049–36064

    PubMed  PubMed Central  Google Scholar 

  • Whelan KA, Caldwell SA, Shahriari KS, Jackson SR, Franchetti LD, Johannes GJ, Reginato MJ (2010) Hypoxia suppression of Bim and Bmf blocks anoikis and luminal clearing during mammary morphogenesis. Mol Biol Cell 21(22):3829–3837

    CAS  PubMed  PubMed Central  Google Scholar 

  • Xing YN, Deng P, Xu HM (2014) Canstatin induces apoptosis in gastric cancer xenograft growth in mice through the mitochondrial apoptotic pathway. Biosci Rep. https://doi.org/10.1042/BSR20140012

  • Xue M, Fang Y, Sun G, Zhuo W, Zhong J, Qian C, Wang L, Wang L, Si J, Chen S (2013) IGFBP3, a transcriptional target of homeobox D10, is correlated with the prognosis of gastric cancer. PLoS ONE 8(12):e81423

    PubMed  PubMed Central  Google Scholar 

  • Yang L, Kuang LG, Zheng HC, Li JY, Wu DY, Zhang SM, Xin Y (2003) PTEN encoding product: a marker for tumorigenesis and progression of gastric carcinoma. World J Gastroenterol 9(1):35–39

    PubMed  PubMed Central  Google Scholar 

  • Zhang Q, Bai X, Chen W, Ma T, Hu Q, Liang C, Xie S, Chen C, Hu L, Xu S, Liang T (2013) Wnt/beta-catenin signaling enhances hypoxia-induced epithelial-mesenchymal transition in hepatocellular carcinoma via crosstalk with hif-1alpha signaling. Carcinogenesis 34(5):962–973

    PubMed  Google Scholar 

  • Zheng XW, Li Y, Tang FA, Ma J, Zheng PY, Lu GF (2009) In vivo antitumor effect of canstatin gene on human esophageal carcinoma xenografts in nude mice. Ai Zheng 28(4):350–355

    CAS  PubMed  Google Scholar 

Download references

Funding

This study was funded by National Research Foundation of Korea (2020R1F1A1073114, 2019R1A2C1090219 and 2014R1A6A9064166) and Korea Institute of Science and Technology Research Project (2V09190).

Author information

Authors and Affiliations

  1. Graduate School of Analytical Science and Technology (GRAST), Chungnam National University, Daejeon, 34134, Republic of Korea

    Seo-Gyu Park, Yoon-Hee Shin, Su-Min Lee & Jae-Young Kim

  2. Advanced Analysis and Data Center, Korea Institute of Science and Technology (KIST), Seoul, 02456, Republic of Korea

    Mi-Jung Ji & Hyun-Mee Park

  3. Department of Surgery, Ajou University School of Medicine, Suwon, 16499, Republic of Korea

    In-Hye Ham & Hoon Hur

  4. Inflammaging Translational Research Cancer, Ajou University School of Medicine, Suwon, 16499, Republic of Korea

    In-Hye Ham & Hoon Hur

  5. Therapeutics and Biotechnology Division, Drug Discovery Platform Research Center, Korea Research Institute of Chemical Technology (KRICT), Daejeon, 34114, Republic of Korea

    Chang Hoon Lee

  6. R and D center, SCBIO Co. Ltd, Daejeon, 34050, Republic of Korea

    Chang Hoon Lee

  7. Natural Product Informatics Center, Korea Institute of Science and Technology (KIST), Gangneung, 25451, Republic of Korea

    Eunjung Kim

  8. Department of Biomedical Science, Graduated School of Ajou University, Suwon, 16499, Republic of Korea

    Hoon Hur

Authors
  1. Seo-Gyu Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mi-Jung Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. In-Hye Ham
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yoon-Hee Shin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Su-Min Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chang Hoon Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Eunjung Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hoon Hur
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hyun-Mee Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Jae-Young Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conception and design: S-GP, J-YK. Acquisition of data: S-GP, M-JJ, I-HH, Y-HS, S-ML. Analysis and interpretation of data: S-GP, CHL, EK, HH, J-YK. Mass spectrometry analysis: M-JJ and H-MP. Writing and reviewing the manuscript: S-GP, EK, HH, EK, J-YK. Study supervision: J-YK. All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Jae-Young Kim.

Ethics declarations

Conflict of Interest

The authors have no conflict of interest to declare.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Rights and permissions

Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Park, SG., Ji, MJ., Ham, IH. et al. Secretome analysis reveals reduced expression of COL4A2 in hypoxic cancer-associated fibroblasts with a tumor-promoting function in gastric cancer. J Cancer Res Clin Oncol 149, 4477–4487 (2023). https://doi.org/10.1007/s00432-022-04361-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00432-022-04361-y

Keywords

Search

Navigation