Ghrelin promotes renal cell carcinoma metastasis via Snail activation and is associated with poor prognosis

• Published in: The Journal of pathology Vol. 237; no. 1; pp. 50 - 61
• Main Authors: Lin, Tsung-Chieh, Liu, Yu-Peng, Chan, Yung-Chieh, Su, Chia-Yi, Lin, Yuan-Feng, Hsu, Shih-Lan, Yang, Chung-Shi, Hsiao, Michael
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.09.2015; Wiley Subscription Services, Inc
• Subjects: Animals; Binding Sites; Cadherins - genetics; Cadherins - metabolism; Carcinoma, Renal Cell - genetics; Carcinoma, Renal Cell - metabolism; Carcinoma, Renal Cell - mortality; Carcinoma, Renal Cell - secondary; Cell Line, Tumor; Cell Movement - drug effects; E-cadherin; Gene Expression Profiling - methods; Gene Expression Regulation, Neoplastic; ghrelin; Ghrelin - genetics; Ghrelin - metabolism; Humans; Kidney Neoplasms - genetics; Kidney Neoplasms - metabolism; Kidney Neoplasms - mortality; Kidney Neoplasms - pathology; Male; metastasis; Mice, Inbred NOD; Mice, SCID; migration; Neoplasm Invasiveness; Oligonucleotide Array Sequence Analysis; Phosphatidylinositol 3-Kinase - antagonists & inhibitors; Phosphatidylinositol 3-Kinase - metabolism; Phosphorylation; Promoter Regions, Genetic - drug effects; Protein Kinase Inhibitors - pharmacology; Proto-Oncogene Proteins c-akt - antagonists & inhibitors; Proto-Oncogene Proteins c-akt - genetics; Proto-Oncogene Proteins c-akt - metabolism; Receptors, Ghrelin - genetics; Receptors, Ghrelin - metabolism; renal cell carcinoma; RNA Interference; Signal Transduction; Snail; Snail Family Transcription Factors; Time Factors; Transcription Factors - genetics; Transcription Factors - metabolism; Transfection; migration; ghrelin; renal cell carcinoma; Snail; metastasis; E-cadherin
• ISSN: 0022-3417; 1096-9896; 1096-9896
• DOI: 10.1002/path.4552

Ghrelin is an appetite‐regulating molecule that promotes growth hormone (GH) release and food intake through growth hormone secretagogue receptor (GHS‐R). Recently, high ghrelin levels have been detected in various types of human cancer. Ghrelin expression is observed in proximal and distal renal tubules, where renal cell carcinoma (RCC) arises. However, whether ghrelin is up‐regulated and promotes renal cell carcinogenesis remains obscure. In this study, we observed that ghrelin was highly expressed in renal tumours, especially in metastatic RCC. In addition, high ghrelin levels correlated with poor outcome, lymph node and distant metastasis. The addition of ghrelin promoted the migration ability of RCC cell lines 786–0, ACHN and A‐498. Furthermore, knockdown of ghrelin expression reduced in vitro migration and in vivo metastasis, suggesting a requirement for ghrelin accumulation in the microenvironment for RCC metastasis. Analysis of microarray signatures using Ingenuity Pathway Analysis (IPA) and MetaCore pointed to the potential regulation by ghrelin of Snail, a transcriptional repressor of E‐cadherin. We further observed that Ghrelin increased the expression, nuclear translocation and promoter‐binding activity of Snail. Snail silencing blocked the ghrelin‐mediated effects on E‐cadherin repression and cell migration. Snail–E‐cadherin regulation was mediated by GHS‐R‐triggered Akt phosphorylation at Ser473 and Thr308. Pretreatment with PI3K inhibitors, LY294002 and wortmannin, as well as Akt siRNA, decreased ghrelin‐induced Akt phosphorylation, Snail promoter binding activity and migration. Taken together, our findings indicate that ghrelin can activate Snail function via the GHS‐R–PI3K–Akt axis, which may contribute to RCC metastasis. The microarray raw data were retrieved from the Cancer Genome Atlas (TCGA) [KIRC gene expression (IlluminaHiSeq) dataset]. Copyright © 2015 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.