Silencing of <em>GRHL2</em> induces epithelial‑to‑mesenchymal transition in lung cancer cell lines with different effects on proliferation and clonogenic growth

Kawabe,Nozomi; Kawabe,Nozomi; Matsuoka,Kohei; Matsuoka,Kohei; Komeda,Kazuki; Komeda,Kazuki; Muraki,Nao; Muraki,Nao; Takaba,Miho; Takaba,Miho; Togami,Yasuha; Togami,Yasuha; Ito,Yumeno; Ito,Yumeno; Yamada,Mizuki; Yamada,Mizuki; Sunaga,Noriaki; Sunaga,Noriaki; Girard,Luc; Girard,Luc; Minna,John D.; Minna,John D.; Cai,Ling; Cai,Ling; Xie,Yang; Xie,Yang; Tanaka,Ichidai; Tanaka,Ichidai; Morise,Masahiro; Morise,Masahiro; Sato,Mitsuo; Sato,Mitsuo

doi:10.3892/ol.2023.13977

Silencing of GRHL2 induces epithelial‑to‑mesenchymal transition in lung cancer cell lines with different effects on proliferation and clonogenic growth

Authors:
- Nozomi Kawabe
- Kohei Matsuoka
- Kazuki Komeda
- Nao Muraki
- Miho Takaba
- Yasuha Togami
- Yumeno Ito
- Mizuki Yamada
- Noriaki Sunaga
- Luc Girard
- John D. Minna
- Ling Cai
- Yang Xie
- Ichidai Tanaka
- Masahiro Morise
- Mitsuo Sato
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Affiliations: Division of Host Defense Sciences, Department of Integrated Health Sciences, Nagoya University Graduate School of Medicine, Nagoya, Aichi 461‑8673, Japan, Department of Respiratory Medicine, Gunma University Graduate School of Medicine, Maebashi, Gunma 371‑8511, Japan, Hamon Center for Therapeutic Oncology Research, Simmons Comprehensive Cancer Center, UT Southwestern Medical Center, Dallas, TX 75230‑8593, USA, Quantitative Biomedical Research Center, Peter O’Donnell School of Public Health, UT Southwestern Medical Center, Dallas, TX 75230‑8593, USA, Department of Respiratory Medicine, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466‑8550, Japan
Published online on: July 25, 2023 https://doi.org/10.3892/ol.2023.13977
Article Number: 391
Copyright: © Kawabe et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Grainyhead‑like 2 (GRHL2) is a transcription factor that suppresses epithelial‑to‑mesenchymal transition (EMT). It has been previously shown that GRHL2 can confer both oncogenic and tumor‑suppressive roles in human cancers, including breast, pancreatic and colorectal cancers. However, its role in lung cancer remains elusive. In the present study, a meta‑analysis of multiple gene expression datasets with clinical data revealed that GRHL2 expression was increased in lung cancer compared with that in the normal tissues. Copy number analysis of GRHL2, performed using datasets of whole exome sequencing involving 151 lung cancer cell lines, revealed frequent amplifications, suggesting that the increased GRHL2 expression may have resulted from gene amplification. A survival meta‑analysis of GRHL2 using The Cancer Genome Atlas (TCGA) dataset showed no association of GRHL2 expression with overall survival. GRHL2 expression was found to be associated with EMT status in lung cancer in TCGA dataset and lung cancer cell lines. GRHL2 knockdown induced partial EMT in the hTERT/Cdk4‑immortalized normal lung epithelial cell line HBEC4KT without affecting proliferation measured by CCK‑8 assays. In addition, GRHL2 silencing caused three lung cancer cell lines, H1975, H2009 and H441, to undergo partial EMT. However, the proliferative effects differed significantly. GRHL2 silencing promoted proliferation but not colony formation in H1975 cells whilst suppressing colony formation without affecting proliferation in H2009 cells, but it did not affect proliferation in H441 cells. These results suggest cell type‑dependent effects of GRHL2 knockdown. Downstream, GRHL2 silencing enhanced the phosphorylation of AKT and ERK, assessed by western blotting with phospho‑specific antibodies, in HBEC4KT, H1975 and H2009 cell lines but not in the H441 cell line. By contrast, transient GRHL2 overexpression did not affect A549 cell proliferation, which lack detectable endogenous expression of the GRHL2 protein. However, GRHL2 overexpression did suppress E‑cadherin expression in A549 cells. These results suggested that GRHL2 does not only function as a tumor suppressor of EMT but can also behave as an oncogene depending on the lung cancer cell‑type context.

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