TFF-1 Functions to Suppress Multiple Phenotypes Associated with Lung Cancer Progression

Kentaro Minegishi; Yoh Dobashi; Hiroyoshi Tsubochi; Koichi Hagiwara; Yuko Ishibashi; Sachiyo Nomura; Ritsuko Nakamura; Yasukazu Ohmoto; Shunsuke Endo

doi:10.2147/OTT.S322697

TFF-1 Functions to Suppress Multiple Phenotypes Associated with Lung Cancer Progression

Onco Targets Ther. 2021 Sep 10:14:4761-4777. doi: 10.2147/OTT.S322697. eCollection 2021.

Authors

Kentaro Minegishi¹, Yoh Dobashi^{2

3}, Hiroyoshi Tsubochi¹, Koichi Hagiwara⁴, Yuko Ishibashi^{5

6}, Sachiyo Nomura⁷, Ritsuko Nakamura⁸, Yasukazu Ohmoto⁹, Shunsuke Endo¹

Affiliations

¹ Department of Thoracic Surgery, Saitama Medical Center, Jichi Medical University, Saitama, Japan.
² Department of Medicine, Saitama Medical Center, Jichi Medical University, Saitama, Japan.
³ Department of Pathology, School of Medicine, International University of Health and Welfare, Tochigi, Japan.
⁴ Division of Pulmonary Medicine, Department of Medicine, Jichi Medical University, Tochigi, Japan.
⁵ Department of Breast and Endocrine Surgery, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
⁶ Division of Breast Surgery, Hospital of the National Center for the Global Health and Medicine, Tokyo, Japan.
⁷ Department of Gastrointestinal Surgery, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
⁸ Department of Molecular and Cellular Pathology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan.
⁹ Tokushima University Industry-University R&D Startup Leading Institute, Tokushima, Japan.

Abstract

Introduction: Trefoil Factor (TFF) is a member of a protein family comprised of three isoforms, of which TFF-1 exhibits antithetical functions; promotion or suppression of cell proliferation, survival and invasion, depending on the cancer type. However, the pathobiological function of TFF-1 in lung carcinoma has been still unclear.

Methods: We examined the expression and secretion of TFF-1 using cultured human lung carcinoma cells by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay and quantitative real-time PCR analyses. The effects of TFF-1 on various phenotypes were analyzed in two cell lines, including those transfected with cDNA encoding TFF-1. Cell proliferation and death were examined by hemocytometer cell counting and by colorimetric viability/cytotoxicity assay. Cell cycle profile, migration and invasion were also examined by flow cytometry, wound healing assay and Matrigel Transwell assay, respectively. The effect of TFF-1 overexpression was confirmed by additional transfection of TFF-1-specific siRNA.

Results: Endogenous TFF-1 protein expression and secretion into the media were observed exclusively in adenocarcinoma-derived cell lines. Forced overexpression of TFF-1 drove cell cycle transition, while the proliferation decreased by 19% to 25% due to increased cell death. This cell death was predominantly caused by apoptosis, as assessed by the activation of caspase 3/7. Cell migration was also suppressed by 71% to 82% in TFF-1-transfected cells. The suppressive effect of TFF-1 on proliferation and migration was restored by transfection of TFF-1 siRNA. Moreover, invasion was also suppressed to 77% to 83% in TFF-1-transfected cells.

Conclusion: These findings reveal that TFF-1 functions as a suppressor of cancer proliferation by induction of apoptosis, cell migration and invasion and thus may provide a synergistic target for potential treatment strategies for human lung carcinoma.

Keywords: TFF-1; apoptosis; growth inhibition; invasion; lung carcinoma cells; migration.