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Lysyl oxidase-like 4 (LOXL4) promotes proliferation and metastasis of gastric cancer via FAK/Src pathway

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

Abstract

Background

Lysyl oxidase-like 4 (LOXL4) has been found up-regulated in a variety of human malignancies, but its clinical significance and functional roles in gastric cancer (GC) remain unknown.

Methods

Lysyl oxidase-like 4 (LOXL4) expression level in tumor tissues and human GC cell lines was evaluated by quantitative real-time polymerase chain reaction, Western blotting and immunohistochemical analyses. Its clinical significance was inferred from the analysis of 379 tissue samples of patients with GC using tissue microarray. The roles of LOXL4 in cell proliferation, migration and invasion in vitro were analyzed by gene over-expression, RNA interference and recombinant protein. Effects of LOXL4 on regulation of focal adhesion kinase/Src kinase (FAK/Src) pathway were examined by Western blotting.

Results

Lysyl oxidase-like 4 (LOXL4) was up-regulated in GC tissues relative to paired non-tumor tissues, and this over-expression was significantly associated with tumor size, depth of tumor invasion, lymph node metastasis, tumor-node-metastasis (TNM) stages and poorer overall survival. Over-expression of LOXL4 has promotive effects on GC cell proliferation, migration and invasion in vitro, consistent with this, LOXL4 knockdown has inhibitive effects on GC cell proliferation, migration and invasion. Furthermore, recombinant human LOXL4 protein also promoted GC cell proliferation and migration. Subsequent mechanistic studies showed that LOXL4 could activate FAK/Src pathway to enhance cell–extracellular matrix adhesion.

Conclusions

Taken together, our data reveal that up-regulation of LOXL4 expression is a frequent event in GC progression, contributes to tumor cell proliferation and metastasis, and LOXL4 may be a potential independent prognostic marker and therapeutic target for GC.

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Abbreviations

ECM:

Extracellular matrix

ERK:

Extracellular regulated kinase

FAK:

Focal adhesion kinase

GC:

Gastric cancer

LOX:

Lysyl oxidase

LOXL4:

Lysyl oxidase-like 4

qRT-PCR:

Quantitative real-time polymerase chain reaction

rhLOXL4:

Recombinant human LOXL4 protein

shRNA:

Short hairpin RNA

TNM:

Tumor node metastasis

References

  • Abe J, Takahashi M, Ishida M, Lee JD, Berk BC (1997) c-Src is required for oxidative stress-mediated activation of big mitogen-activated protein kinase 1. J Biol Chem 272:20389–20394

    Article  CAS  PubMed  Google Scholar 

  • Bae YS, Kang SW, Seo MS, Baines IC, Tekle E, Chock PB, Rhee SG (1997) Epidermal growth factor (EGF)-induced generation of hydrogen peroxide. Role in EGF receptor-mediated tyrosine phosphorylation. J Biol Chem 272:217–221

    Article  CAS  PubMed  Google Scholar 

  • Baker AM, Cox TR, Bird D, Lang G, Murray GI, Sun XF, Southall SM, Wilson JR, Erler JT (2011) The role of lysyl oxidase in SRC-dependent proliferation and metastasis of colorectal cancer. Natl Cancer Inst 103:407–424

    Article  CAS  Google Scholar 

  • Baker AM, Bird D, Lang G, Cox TR, Erler JT (2013) Lysyl oxidase enzymatic function increases stiffness to drive colorectal cancer progression through FAK. Oncogene 32:1863–1868

    Article  CAS  PubMed  Google Scholar 

  • Crew KD, Neugut AI (2006) Epidemiology of gastric cancer. World J Gastroenterol 12:354–362

    PubMed Central  PubMed  Google Scholar 

  • Csiszar K (2001) Lysyl oxidases: a novel multifunctional amine oxidase family. Prog Nucleic Acid Res Mol Biol 70:1–32

    Article  CAS  PubMed  Google Scholar 

  • Görögh T, Weise JB, Holtmeier C, Rudolph P, Hedderich J, Gottschlich S, Hoffmann M, Ambrosch P, Csiszar K (2007) Selective upregulation and amplification of the lysyl oxidase like-4 (LOXL4) gene in head and neck squamous cell carcinoma. J Pathol 212:74–82

    Article  PubMed  Google Scholar 

  • Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D (2011) Global cancer statistics. CA Cancer J Clin 61:69–90

    Article  PubMed  Google Scholar 

  • Jones RJ, Brunton VG, Frame MC (2000) Adhesion-linked kinases in cancer; emphasis on src, focal adhesion kinase and PI 3-kinase. Eur J Cancer 36:1595–1606

    Article  CAS  PubMed  Google Scholar 

  • Kagan HM, Li W (2003) Lysyl Oxidase: properties, specificity, and biological roles inside and outside of the cell. J Cell Biochem 88:660–672

    Article  CAS  PubMed  Google Scholar 

  • Kaplan KB, Swedlow JR, Morgan DO, Varmus HE (1995) c-Src enhances the spreading of src−/− fibroblasts on fibronectin by a kinase-independent mechanism. Genes Dev 9:1505–1517

    Article  CAS  PubMed  Google Scholar 

  • Kim Y, Roh S, Park JY, Kim Y, Cho DH, Kim JC (2009) Differential expression of the LOX family genes in human colorectal adenocarcinomas. Oncol Rep 22:799–804

    CAS  PubMed  Google Scholar 

  • Kirschmann DA, Seftor EA, Fong SF, Nieva DR, Sullivan CM, Edwards EM, Sommer P, Csiszar K, Hendrix MJ (2002) A molecular role for lysyl oxidase in breast cancer invasion. Cancer Res 62:4478–4483

    CAS  PubMed  Google Scholar 

  • Li J, Yang X-M, Wang Y-H, Feng M-X, Liu X-J, Zhang Y-L, Huang S, Wu Z, Xue F, Qin W-X, Gu J-R, Xia Q, Zhang Z-G (2014) Monoamine oxidase A suppresses hepatocellular carcinoma metastasis by inhibiting the adrenergic system and its transactivation of EGFR signaling. J Hepatol 60:1225–1234

    Article  CAS  PubMed  Google Scholar 

  • Ma MZ, Zhuang C, Yang XM, Zhang ZZ, Ma H, Zhang WM, You HY, Qin W, Gu J, Yang S, Cao H, Zhang ZG (2014) CTHRC1 acts as a prognostic factor and promotes invasiveness of gastrointestinal stromal tumors by activating Wnt/PCP-Rho signaling. Neoplasia 16:265–278

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Mäki JM, Tikkanen H, Kivirikko KI (2001) Cloning and characterization of a fifth human lysyl oxidase isoenzyme: the third member of the lysyl oxidase-related subfamily with four scavenger receptor cysteine-rich domains. Matrix Biol 20:493–496

    Article  PubMed  Google Scholar 

  • Manes S, Mira E, Gomez-Mouton C, Lacalle RA, Martinez C (2000) Cells on the move: a dialogue between polarization and motility. IUBMB Life 49:89–96

    Article  CAS  PubMed  Google Scholar 

  • Maung K, Easty DJ, Hill SP, Bennett DC (1999) Requirement for focal adhesion kinase in tumor cell adhesion. Oncogene 18:6824–6828

    Article  CAS  PubMed  Google Scholar 

  • Nada S, Yagi T, Takeda H, Tokunaga T, Nakagawa H, Ikawa Y, Okada M, Aizawa S (1993) Constitutive activation of Src family kinases in mouse embryos that lack Csk. Cell 73:1125–1135

    Article  CAS  PubMed  Google Scholar 

  • Naylor GM, Gotoda T, Dixon M, Shimoda T, Gatta L, Owen R, Tompkins D, Axon A (2006) Why does Japan have a high incidence of gastric cancer? Comparison of gastritis between UK and Japanese patients. Gut 55:1545–1552

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Payne SL, Fogelgren B, Hess AR, Seftor EA, Wiley EL, Fong SF, Csiszar K, Hendrix MJ, Kirschmann DA (2005) Lysyl oxidase regulates breast cancer cell migration and adhesion through a hydrogen peroxide-mediated mechanism. Cancer Res 65:11429–11436

    Article  CAS  PubMed  Google Scholar 

  • Payne SL, Hendrix MJ, Kirschmann DA (2007) Paradoxical roles for lysyl oxidases in cancer—a prospect. J Cell Biochem 101:1338–1354

    Article  CAS  PubMed  Google Scholar 

  • Peng L, Ran Y-L, Hai H, Yu L, Liu Q, Zhou Z, Sun Y-M, Sun L-C, Pan J, Sun L-X, Zhao P, Yang Z-H (2009) Secreted LOXL2 is a novel therapeutic target that promotes gastric cancer metastasis via the Src/FAK pathway. Carcinogenesis 30:1660–1669

    Article  CAS  PubMed  Google Scholar 

  • Schlaepfer DD, Jones KC, Hunter T (1998) Multiple Grb2-mediated integrin-stimulated signaling pathways to ERK2/mitogen-activated protein kinase: summation of both c-Src and focal adhesion kinase-initiated tyrosine phosphorylation events. Mol Cell Biol 18:2571–2585

    CAS  PubMed Central  PubMed  Google Scholar 

  • Stracke ML, Liotta LA (1992) Multi-step cascade of tumor cell metastasis. In Vivo 6:309–316

    CAS  PubMed  Google Scholar 

  • Sun W, Haller DG (2001) Recent advances in the treatment of gastric cancer. Drugs 61:1545–1551

    Article  CAS  PubMed  Google Scholar 

  • Weise JB, Rudolph P, Heiser A, Kruse ML, Hedderich J, Cordes C, Hoffmann M, Brant O, Ambrosch P, Csiszar K, Görögh T (2008) LOXL4 is a selectively expressed candidate diagnostic antigen in head and neck cancer. Eur J Cancer 44:1323–1331

    Article  CAS  PubMed  Google Scholar 

  • Wong CC, Zhang H, Gilkes DM, Chen J, Wei H, Chaturvedi P, Hubbi ME, Semenza GL (2012) Inhibitors of hypoxia-inducible factor 1 block breast cancer metastatic niche formation and lung metastasis. J Mol Med (Berl) 90:803–815

    Article  CAS  Google Scholar 

  • Wu G, Guo Z, Chang X, Kim MS, Nagpal JK, Liu J, Maki JM, Kivirikko KI, Ethier SP, Trink B, Sidransky D (2007) LOXL1 and LOXL4 are epigenetically silenced and can inhibit Ras/extracellular signal-regulated kinase signaling pathway in human bladder cancer. Cancer Res 67:4123–4129

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We would like to thank the stuff of the tumor microenvironment research group of State Key Laboratory of Oncogenes and Related Genes for assistance with this study.

Conflict of interest

The authors have declared no conflict of interest.

Ethical standard

This project was approved by committee of Renji Hospital, Shanghai Jiaotong University School of Medicine for the use of samples. All tissue samples were obtained with the consent of patients and in accordance with the China Ethical Review Committee.

Author information

Authors and Affiliations

  1. State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, 800 Dongchuan Road, Shanghai, 200240, China

    Rong-kun Li, Fang Fang, Xiao-xin Zhang, Xiao-mei Yang, Shu-heng Jiang, Sheng-li Yang & Zhi-gang Zhang

  2. Department of Oncology, Huai’an First People’s Hospital, Nanjing Medical University, Huai’an, 223300, China

    Rong-kun Li

  3. Department of General Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, 1630 Dongfang Road, Shanghai, 200127, China

    Wen-yi Zhao, Chun Zhuang, Lin Tu & Hui Cao

  4. Department of General Surgery, Central Hospital of Fengxian District, Shanghai, 201400, China

    Fan-zhi Kong

  5. Department of Endoscopy, Cancer Hospital, Fudan University, Shanghai, 200032, China

    Wen-Ming Zhang

  6. Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, 200032, China

    Wen-Ming Zhang

Authors
  1. Rong-kun Li
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  2. Wen-yi Zhao
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  3. Fang Fang
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  9. Lin Tu
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  11. Sheng-li Yang
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  12. Hui Cao
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  13. Zhi-gang Zhang
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Corresponding authors

Correspondence to Hui Cao or Zhi-gang Zhang.

Additional information

Rong-kun Li and Wen-yi Zhao have contributed equally to this work.

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Li, Rk., Zhao, Wy., Fang, F. et al. Lysyl oxidase-like 4 (LOXL4) promotes proliferation and metastasis of gastric cancer via FAK/Src pathway. J Cancer Res Clin Oncol 141, 269–281 (2015). https://doi.org/10.1007/s00432-014-1823-z

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  • DOI: https://doi.org/10.1007/s00432-014-1823-z

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