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Carcinogenesis Advance Access published online on February 4, 2004
Carcinogenesis, doi:10.1093/carcin/bgh097
© 2004 by Oxford University Press
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© 2004 Oxford University Press

CANCER BIOLOGY

Overexpression of gastrin-releasing peptide in human esophageal squamous cell carcinomas

Ming Zhu Fang 1, Changgong Liu 1, Yunlong Song 1, Guang-Yu Yang 1, Yan Nie 1, Jie Liao 1, Xin Zhao 2, Yutaka Shimada 3, Li-Dong Wang 2, and Chung S. Yang 1*

1 Susan Lehman Cullman Laboratory for Cancer Research, Department of Chemical Biology, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, NJ 08854-8020, USA
2 Laboratory for Cancer Research, College of Medicine, Zhengzhou University, Zhengzhou, Henan 450052, China
3 Department of Surgery and Surgical Basic Science, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan

* Corresponding author. E-mail: csyang{at}rci.rutgers.edu.

Received 27 May 2003 ; revised 12 December 2003 ; accepted 13 January 2004

Abstract

Gastrin-releasing peptide (GRP) is known as an autocrine growth factor for a number of gastrointestinal cancers. There is, however, little information on the expression of GRP in the squamous epithelia and squamous cell carcinoma, particularly in the esophagus. With a differential display approach, up-regulated GRP was observed in human ESCC samples obtained from a high-risk area for esophageal cancer, Linzhou in northern China. Up-regulation of phosphoglycerate mutase and P311 HUM (3.1) and down-regulation of keratin 13, cystatin B, endoglin, and annexin I were observed. Using a reverse transcription-polymerase chain reaction (RT-PCR) method, significant overexpression of GRP was observed in 10 out of 12 ESCC samples (83.3%) and all 4 ESCC cell lines. With in situ hybridization, GRP mRNA expression was detected in 9 out of 21 (42.8%) samples with basal cell hyperplasia (BCH), 5 out of 7 (71.4%) samples with dysplasia (DYS), and 17 out of 24 (70.9%) ESCC samples. In contrast, GRP was expressed only in 3 out of 16 (18.7%) normal epithelium. Digital image analysis revealed that the mean value of GRP expression index, determined by intensity and area ratio of staining, was 0.19 in normal epithelium, 1.23 in BCH, 2.94 in DYS, and 2.38 in ESCC, showing a progressive increase. Studies on ESCC cell lines showed GRP increased cell growth in a dose dependent pattern in GRP receptor-positive ESCC cells, but not in GRP receptor-negative ESCC cells. GRP (1 µM) also increased cyclooxygenase-2 protein expression by 3.4 fold. This is the first demonstration that GRP is overexpressed in ESCC, and its overexpression may play a role in ESCC development and growth.

gastrin-releasing peptide, esophageal squamous cell carcinoma, in situ hybridization, cell proliferation
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