Tao Xie1, Yifan Tang1, Rongkui Luo2, Xiaobiao Zhang3,4,5, Silin Wu1, Ye Gu1, Tengfei Liu1, Fan Hu1. 1. Department of Neurosurgery, Zhongshan Hospital, Fudan University, Shanghai, China. 2. Department of Pathology, Zhongshan Hospital, Fudan University, Shanghai, China. 3. Department of Neurosurgery, Zhongshan Hospital, Fudan University, Shanghai, China. xiaobiao_zhang@163.com. 4. Department of Digital Medical Research Center, Fudan University, Shanghai, China. xiaobiao_zhang@163.com. 5. Shanghai Key Laboratory of Medical Image Computing and Computer-Assisted Intervention, Shanghai, China. xiaobiao_zhang@163.com.
Abstract
PURPOSE: There is an increasing agreement that acromegaly caused by growth hormone (GH) cell adenoma has two distinct subtypes: densely granulated (DG) and sparsely granulated (SG). We hypothesized that differential molecular signatures may explain their behavior. METHODS: Total transcriptome sequencing was performed on ten DG and seven SG adenomas. The differentially expressed RNAs were identified by bioinformatic analyses, and a candidate RNA was verified by quantitative real-time PCR. Immunohistochemical staining was also performed to detect the protein expression of the candidate. Clinical parameters were correlated with protein expression. Subsequently, cell proliferation, colony formation, and cell cycle progression were analyzed after knockdown of the candidate in pituitary GH3 cells. Activation of the cAMP pathway was assessed by ELISA and Western blot. RESULTS: We confirmed that there were obvious differentially expressed genes between the subtypes. Through gene profiling, we discovered that an orphan adhesion G protein-coupled receptor, GPR64, was overexpressed in more aggressive SG adenomas. Noticeably, GPR64 knockdown significantly inhibited the proliferation of GH3 tumor cells and decreased colony formation. The knockdown also induced cell cycle arrest in GH3 tumor cells. Further studies revealed that GPR64 knockdown decreased cAMP levels and the ratios of p-CREB/CREB, indicating that it suppressed the cAMP/CREB pathway. CONCLUSIONS: Our results indicated that GPR64 may promote aggressiveness in SG-type GH cell adenomas and that it is a key factor regulating the cAMP pathway to promote aggressiveness of GH cell adenomas.
PURPOSE: There is an increasing agreement that acromegaly caused by growth hormone (GH) cell adenoma has two distinct subtypes: densely granulated (DG) and sparsely granulated (SG). We hypothesized that differential molecular signatures may explain their behavior. METHODS: Total transcriptome sequencing was performed on ten DG and seven SG adenomas. The differentially expressed RNAs were identified by bioinformatic analyses, and a candidate RNA was verified by quantitative real-time PCR. Immunohistochemical staining was also performed to detect the protein expression of the candidate. Clinical parameters were correlated with protein expression. Subsequently, cell proliferation, colony formation, and cell cycle progression were analyzed after knockdown of the candidate in pituitary GH3 cells. Activation of the cAMP pathway was assessed by ELISA and Western blot. RESULTS: We confirmed that there were obvious differentially expressed genes between the subtypes. Through gene profiling, we discovered that an orphan adhesion G protein-coupled receptor, GPR64, was overexpressed in more aggressive SG adenomas. Noticeably, GPR64 knockdown significantly inhibited the proliferation of GH3 tumor cells and decreased colony formation. The knockdown also induced cell cycle arrest in GH3 tumor cells. Further studies revealed that GPR64 knockdown decreased cAMP levels and the ratios of p-CREB/CREB, indicating that it suppressed the cAMP/CREB pathway. CONCLUSIONS: Our results indicated that GPR64 may promote aggressiveness in SG-type GH cell adenomas and that it is a key factor regulating the cAMP pathway to promote aggressiveness of GH cell adenomas.
Authors: Katja Kiseljak-Vassiliades; Nichole E Carlson; Manuel T Borges; B K Kleinschmidt-DeMasters; Kevin O Lillehei; Janice M Kerr; Margaret E Wierman Journal: Endocrine Date: 2014-08-17 Impact factor: 3.633
Authors: Iulia Potorac; Patrick Petrossians; Adrian F Daly; Orsalia Alexopoulou; Sophie Borot; Mona Sahnoun-Fathallah; Frederic Castinetti; France Devuyst; Marie-Lise Jaffrain-Rea; Claire Briet; Florina Luca; Marion Lapoirie; Flavius Zoicas; Isabelle Simoneau; Alpha M Diallo; Ammar Muhammad; Fahrettin Kelestimur; Elena Nazzari; Rogelio Garcia Centeno; Susan M Webb; Marie-Laure Nunes; Vaclav Hana; Véronique Pascal-Vigneron; Irena Ilovayskaya; Farida Nasybullina; Samia Achir; Diego Ferone; Sebastian J C M M Neggers; Brigitte Delemer; Jean-Michel Petit; Christof Schöfl; Gerald Raverot; Bernard Goichot; Patrice Rodien; Bernard Corvilain; Thierry Brue; Franck Schillo; Luaba Tshibanda; Dominique Maiter; Jean-François Bonneville; Albert Beckers Journal: Endocr Relat Cancer Date: 2016-09-20 Impact factor: 5.678