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Ezrin-expressing lung adenocarcinoma cells and podoplanin-positive fibroblasts form a malignant microenvironment

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

Abstract

Purpose

Cancer cells and cancer-associated fibroblasts (CAFs) together create the tumor microenvironment, which affects malignant behavior. Lung adenocarcinomas with CAFs expressing podoplanin (PDPN) are clinically aggressive, but the molecular mechanism underlying this phenomenon has not been established. So we identified the characteristic immunophenotype of lung adenocarcinoma cells coexisting with PDPN-expressing CAFs (PDPN-CAFs) and examined how it relates to an aggressive clinicopathological outcome.

Methods

We analyzed the clinicopathological characteristics of 119 adenocarcinomas with a uniform size (2–3 cm). The expression levels of ten invasiveness-related proteins which related to cell adhesion and invasiveness, such as Ezrin, were examined in cancer cells from PDPN-CAFs (+) cases and from PDPN-CAFs (−) cases (n = 20 each). To examine the functional importance of the identified protein on the invasion phenotype, we performed wound healing and a Matrigel invasion assay using shRNA-knockdown lung adenocarcinoma cells (PC-9).

Results

The PDPN-CAFs (+) cases had significantly higher rates of node metastasis (p < 0.01) and vascular invasion (p < 0.01). The cancer cells from the PDPN-CAFs (+) cases also had a significantly higher staining score for Ezrin (p < 0.01) than those from the PDPN-CAFs (−) cases. The migration and invasion activities of the shEzrin-induced PC-9 cells were significantly lower than those of the control cells.

Conclusions

Our results indicated that within a tumor microenvironment composed of PDPN-CAFs, increased Ezrin expression in cancer cells might play a key role in the invasiveness of lung adenocarcinoma.

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Acknowledgments

This work was supported by National Cancer Center Research and Development Fund (23-A-12 and 23-K-18), the Foundation for the Promotion of Cancer Research, 3rd-Term Comprehensive 10-Year Strategy for Cancer Control, the Advanced research for medical products Mining Programme of the National Institute of Biomedical Innovation (NIBIO), and JSPS KAKENHI (24659185).

Conflict of interest

The authors have no conflict of interest to disclose.

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Authors and Affiliations

  1. Pathology Division, Research Center for Innovative Oncology, National Cancer Center Hospital East, Kashiwanoha, Kashiwa, Chiba, 277-8577, Japan

    Shigeki Suzuki, Genichiro Ishii, Rie Matsuwaki, Shinya Neri, Hiroko Hashimoto, Chisako Yamauchi & Atsushi Ochiai

  2. Division of Thoracic Surgery, National Cancer Center Hospital East, Kashiwa, Chiba, Japan

    Shigeki Suzuki, Rie Matsuwaki, Shinya Neri, Keiju Aokage, Tomoyuki Hishida, Junji Yoshida & Kanji Nagai

  3. Division of General Thoracic Surgery, Department of Surgery, Keio University School of Medicine, Tokyo, Japan

    Shigeki Suzuki & Mitsutomo Kohno

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  1. Shigeki Suzuki
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  2. Genichiro Ishii
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  3. Rie Matsuwaki
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Correspondence to Genichiro Ishii.

Electronic supplementary material

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432_2014_1851_MOESM1_ESM.pdf

Supplementary Figure 1 Western blot analysis showing Ezrin expression in control (shLuciferase) or Ezrin shRNA-induced PC-9 cells, compared with β-actin expression (PDF 71 kb)

Supplementary material 2 (DOCX 18 kb)

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Supplementary material 4 (DOCX 20 kb)

Supplementary material 5 (DOCX 18 kb)

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Suzuki, S., Ishii, G., Matsuwaki, R. et al. Ezrin-expressing lung adenocarcinoma cells and podoplanin-positive fibroblasts form a malignant microenvironment. J Cancer Res Clin Oncol 141, 475–484 (2015). https://doi.org/10.1007/s00432-014-1851-8

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  • DOI: https://doi.org/10.1007/s00432-014-1851-8

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