Bone morphogenetic protein 4 provides cancer-supportive phenotypes to liver fibroblasts in patients with hepatocellular carcinoma

  • Yohei Mano
  • Sachiyo Yoshio
  • Hirotaka Shoji
  • Shimagaki Tomonari
  • Yoshihiko Aoki
  • Nobuyoshi Aoyanagi
  • Toru Okamoto
  • Yoshiharu Matsuura
  • Yosuke Osawa
  • Kiminori Kimura
  • Kyohei Yugawa
  • Huanlin Wang
  • Yoshinao Oda
  • Tomoharu Yoshizumi
  • Yoshihiko Maehara
  • Tatsuya KantoEmail author
Original Article—Liver, Pancreas, and Biliary Tract



Cancer-associated fibroblasts (CAFs) are essential constituents of cancer-supportive microenvironments. The high incidence of hepatocellular carcinoma (HCC) in advanced fibrosis patients implies that fibroblasts have a promoting effect on HCC development. We aimed to explore the regulators of phenotypes and function of CAFs in the liver.


We established primary cancer-associated fibroblasts (CAFs) and non-cancerous liver fibroblasts (NFs) from 15 patients who underwent HCC resection. We compared phenotypes, capacity of cytokine/chemokine production and gene expression profiles between pairs of CAFs and NFs from the same donors. We examined resected tissue from additional 50 patients with HCC for immunohistochemical analyses.


The CAFs expressed more ACTA2 and COL1A1 than the NFs, suggesting that CAFs are more activated phenotype. The CAFs produced larger amounts of IL-6, IL-8 and CCL2 than the NFs, which led to invasiveness of HuH7 in vitro. We found that Bone Morphogenetic Protein-4 (BMP4) is up-regulated in CAFs compared to NFs. The CAF phenotype and function were gained by BMP4 over-expression or recombinant BMP4 given to fibroblasts, all of which decreased with BMP4 knockdown. In tissues obtained from the patients, BMP4-positive cells are mainly observed in encapsulated fibrous lesions and HCC. Positive expression of BMP4 in HCC in resected tissues, not in fibroblasts, was associated with poorer postoperative overall survival in patients with HCC.


Endogenous and exogenous BMP4 activate liver fibroblasts to gain capacity of secreting cytokines and enhancing invasiveness of cancer cells in the liver. BMP4 is one of the regulatory factors of CAFs functioning in the microenvironment of HCC.


Cancer-associated fibroblasts Tumor microenvironment IL-6 IL-8 



Bone morphogenetic protein-4


Cancer-associated fibroblasts


Conditioned media


Direct anti-viral agents


Hepatocellular carcinoma


Hepatic stellate cells


Non-cancerous liver fibroblasts


Overall survival



We thank Ms. Chizu Kanokoda for her technical assistance.


This study was supported by Grants-in-aid for Research from the National Center for Global Health and Medicine (26-shi-109 and 26A201) and by the Research Program on Hepatitis from Japan Agency for Medical Research and Development (17fk0210305h0003).

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Supplementary material

535_2019_1579_MOESM1_ESM.docx (1.9 mb)
Supplementary material 1 (DOCX 1939 kb)


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Copyright information

© Japanese Society of Gastroenterology 2019

Authors and Affiliations

  • Yohei Mano
    • 1
    • 2
  • Sachiyo Yoshio
    • 1
  • Hirotaka Shoji
    • 1
  • Shimagaki Tomonari
    • 1
    • 2
  • Yoshihiko Aoki
    • 1
  • Nobuyoshi Aoyanagi
    • 3
  • Toru Okamoto
    • 4
  • Yoshiharu Matsuura
    • 4
  • Yosuke Osawa
    • 1
  • Kiminori Kimura
    • 5
  • Kyohei Yugawa
    • 2
    • 6
  • Huanlin Wang
    • 2
    • 6
  • Yoshinao Oda
    • 6
  • Tomoharu Yoshizumi
    • 2
  • Yoshihiko Maehara
    • 7
  • Tatsuya Kanto
    • 1
    Email author
  1. 1.The Research Center for Hepatitis and ImmunologyNational Center for Global Health and MedicineIchikawaJapan
  2. 2.Department of Surgery and Science, Graduate School of Medical SciencesKyushu UniversityFukuokaJapan
  3. 3.Department of Surgery, Kohnodai HospitalNational Center for Global Health and MedicineIchikawaJapan
  4. 4.Department of Molecular Virology, Research Institute for Microbial DiseasesOsaka UniversitySuitaJapan
  5. 5.Department of HepatologyTokyo Metropolitan Cancer and Infectious Diseases Center Komagome HospitalTokyoJapan
  6. 6.Department of Anatomic Pathology, Graduate School of Medical SciencesKyushu UniversityFukuokaJapan
  7. 7.Department of SurgeryKyushu Central HospitalFukuokaJapan

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