Forced Trefoil Factor Family Peptide 3 (TFF3) Expression Reduces Growth, Viability, and Tumorigenicity of Human Retinoblastoma Cell Lines.
Trefoil factor family (TFF) peptides have been shown to effect cell proliferation, apoptosis, migration and invasion of normal cells and various cancer cell lines. In the literature TFF peptides are controversially discussed as tumor suppressors and potential tumor progression factors. In the study presented, we investigated the effect of TFF3 overexpression on growth, viability, migration and tumorigenicity of the human retinoblastoma cell lines Y-79, WERI-Rb1, RBL-13 and RBL-15. As revealed by WST-1 and TUNEL assays as well as DAPI and BrdU cell counts, recombinant human TFF3 significantly lowers retinoblastoma cell viability and increases apoptosis levels. Transient TFF3 overexpression likewise significantly increases RB cell apoptosis. Stable, lentiviral TFF3 overexpression lowers retinoblastoma cell viability, proliferation and growth and significantly increases cell death in retinoblastoma cells. Blockage experiments using a broad-spectrum caspase inhibitor and capase-3 immunocytochemistry revealed the involvement of caspases in general and of caspase-3 in particular in TFF3 induced apoptosis in retinoblastoma cell lines. Soft agarose and in ovo chicken chorioallantoic membrane (CAM) assays revealed that TFF3 overexpression influences anchorage independent growth and significantly decreases the size of tumors forming from retinoblastoma cells. Our study demonstrates that forced TFF3 expression exerts a significant pro-apoptotic, anti-proliferative, and tumor suppressive effect in retinoblastoma cells, setting a starting point for new additive chemotherapeutic approaches in the treatment of retinoblastoma.
University of Duisburg-Essen, Medical Faculty, Institute of Anatomy II, Department of Neuroanatomy, Essen, Germany.
Jan
J
Große-Kreul
University of Duisburg-Essen, Medical Faculty, Institute of Anatomy II, Department of Neuroanatomy, Essen, Germany.
Maike
M
Busch
University of Duisburg-Essen, Medical Faculty, Institute of Anatomy II, Department of Neuroanatomy, Essen, Germany.
Claudia
C
Winter
University of Duisburg-Essen, Medical Faculty, Institute of Anatomy II, Department of Neuroanatomy, Essen, Germany.
Stefanie
S
Pikos
Division of Haematology and Oncology, Children's Hospital, University of Duisburg-Essen, Essen, Germany.
Harald
H
Stephan
University of Duisburg-Essen, Medical Faculty, Institute of Anatomy II, Department of Neuroanatomy, Essen, Germany.
Nicole
N
Dünker
D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D019572:Retinal Neoplasms / Q000378:metabolism* / Q000503:physiopathology; D012175:Retinoblastoma / Q000378:metabolism* / Q000503:physiopathology; D000071165:Trefoil Factor-3 / Q000378:metabolism / Q000502:physiology*
C082495:TFF3 protein, human; D000071165:Trefoil Factor-3