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Molecular Cancer
Published in: Molecular Cancer 1/2014

Open Access 01-12-2014 | Research

EGFR phosphorylates and inhibits lung tumor suppressor GPRC5A in lung cancer

Authors: Xiaofeng Lin, Shuangshuang Zhong, Xiaofeng Ye, Yueling Liao, Feng Yao, Xiaohua Yang, Beibei Sun, Jie Zhang, Qi Li, Yong Gao, Yifan Wang, Jingyi Liu, Baohui Han, Y Eugene Chin, Binhua P Zhou, Jiong Deng

Published in: Molecular Cancer | Issue 1/2014

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Abstract

Background

GPRC5A is a retinoic acid inducible gene that is preferentially expressed in lung tissue. Gprc5a– knockout mice develop spontaneous lung cancer, indicating Gprc5a is a lung tumor suppressor gene. GPRC5A expression is frequently suppressed in majority of non-small cell lung cancers (NSCLCs), however, elevated GPRC5A is still observed in a small portion of NSCLC cell lines and tumors, suggesting that the tumor suppressive function of GPRC5A is inhibited in these tumors by an unknown mechanism.

Methods

In this study, we examined EGF receptor (EGFR)-mediated interaction and tyrosine phosphorylation of GPRC5A by immunoprecipitation (IP)-Westernblot. Tyrosine phosphorylation of GPRC5A by EGFR was systematically identified by site-directed mutagenesis. Cell proliferation, migration, and anchorage-independent growth of NSCLC cell lines stably transfected with wild-type GPRC5A and mutants defective in tyrosine phosphorylation were assayed. Immunohistochemical (IHC) staining analysis with specific antibodies was performed to measure the total and phosphorylated GPRC5A in both normal lung and lung tumor tissues.

Result

We found that EGFR interacted with GPRC5A and phosphorylated it in two conserved double-tyrosine motifs, Y317/Y320 and Y347/ Y350, at the C-terminal tail of GPRC5A. EGF induced phosphorylation of GPRC5A, which disrupted GPRC5A-mediated suppression on anchorage-independent growth of NSCLC cells. On contrary, GPRC5A-4 F, in which the four tyrosine residues have been replaced with phenylalanine, was resistant to EGF-induced phosphorylation and maintained tumor suppressive activities. Importantly, IHC analysis with anti-Y317/Y320-P sites showed that GPRC5A was non-phosphorylated in normal lung tissue whereas it was highly tyrosine-phosphorylated in NSCLC tissues.

Conclusion

GPRC5A can be inactivated by receptor tyrosine kinase via tyrosine phosphorylation. Thus, targeting EGFR can restore the tumor suppressive functions of GPRC5A in lung cancer.
Appendix
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Metadata
Title
EGFR phosphorylates and inhibits lung tumor suppressor GPRC5A in lung cancer
Authors
Xiaofeng Lin
Shuangshuang Zhong
Xiaofeng Ye
Yueling Liao
Feng Yao
Xiaohua Yang
Beibei Sun
Jie Zhang
Qi Li
Yong Gao
Yifan Wang
Jingyi Liu
Baohui Han
Y Eugene Chin
Binhua P Zhou
Jiong Deng
Publication date
01-12-2014
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2014
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/1476-4598-13-233

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