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Article: Differential expression of human gonadotropin-releasing hormone receptor gene in pituitary and ovarian cells

TitleDifferential expression of human gonadotropin-releasing hormone receptor gene in pituitary and ovarian cells
Authors
KeywordsDifferential expression
Human GnRH receptor
Ovary
Transcription factors
Issue Date2000
PublisherElsevier Ireland Ltd. The Journal's web site is located at http://www.elsevier.com/locate/mce
Citation
Molecular And Cellular Endocrinology, 2000, v. 162 n. 1-2, p. 157-166 How to Cite?
AbstractIn terms of regulation of gene expression, gonadotropin-releasing hormone receptor (GnRHR) found in extrapituitary tissues has been suggested to be different from that in the pituitary. In the present study, we examined the molecular basis of this difference using the pituitary αT3-1 and ovarian carcinoma OVCAR-3 cells. As a first step, the different expression levels of GnRHR mRNA in the pituitary and ovarian cells were investigated using semi- quantitative RT-PCR. Quantitative analysis showed that the expression level of hGnRHR is a nine-fold higher in primary pituitary tissues than the primary culture of ovarian carcinomas (PCO). In pituitary αT3-1 cells, the expression level of hGnRHR was ten-fold higher than ovarian carcinoma OVCAR-3 cells. The possibility of the differential use of various cell-specific promoters in different cells was addressed by transiently transfecting cells with 3'-deletion clones of human GnRHR promoter. Sequential deletion of the 5'-flanking region of the gene revealed the use of two putative promoters, designated PR1 (-771 to -557) and PR2 (-1351 to -1022), and a negative control region (-1022 to -771), in the pituitary and ovarian cells. The same promoters appeared to be utilized for driving the basal promoter activities in both αT3-1 and OVCAR-3 cells, suggesting that there is no cell-specific promoter usage for the human GnRHR gene. Alternatively, the involvement of different regulatory protein factors was investigated using electrophoretic gel mobility shift assays. When end-labeled PR1 was used as a probe, two unique shifted complexes were identified in OVCAR-3 cells when compared to αT3-1 cells. One unique protein-DNA complex was observed in αT3-1 cells compared to OVCAR-3 cells when incubated with end-labeled PR2 as a probe. These DNA-protein complexes appeared to be specific, as they competed with excess amount of unlabelled competitor PR1 and PR2, respectively. In summary, it is unlikely that the use of a cell-specific promoter contributes to the different characteristics of ovarian GnRHR. Our study demonstrates that one mechanism by which cell-specific expression of human GnRHR is achieved is through the binding of distinct and/or combinations of cell-specific regulatory factors to various promoter elements in the 5'-flanking region of the gene. (C) 2000 Elsevier Science Ireland Ltd.
Persistent Identifierhttp://hdl.handle.net/10722/84906
ISSN
2015 Impact Factor: 3.859
2015 SCImago Journal Rankings: 2.116
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorKang, SKen_HK
dc.contributor.authorCheng, KWen_HK
dc.contributor.authorNgan, ESWen_HK
dc.contributor.authorChow, BKCen_HK
dc.contributor.authorChoi, KCen_HK
dc.contributor.authorLeung, PCKen_HK
dc.date.accessioned2010-09-06T08:58:30Z-
dc.date.available2010-09-06T08:58:30Z-
dc.date.issued2000en_HK
dc.identifier.citationMolecular And Cellular Endocrinology, 2000, v. 162 n. 1-2, p. 157-166en_HK
dc.identifier.issn0303-7207en_HK
dc.identifier.urihttp://hdl.handle.net/10722/84906-
dc.description.abstractIn terms of regulation of gene expression, gonadotropin-releasing hormone receptor (GnRHR) found in extrapituitary tissues has been suggested to be different from that in the pituitary. In the present study, we examined the molecular basis of this difference using the pituitary αT3-1 and ovarian carcinoma OVCAR-3 cells. As a first step, the different expression levels of GnRHR mRNA in the pituitary and ovarian cells were investigated using semi- quantitative RT-PCR. Quantitative analysis showed that the expression level of hGnRHR is a nine-fold higher in primary pituitary tissues than the primary culture of ovarian carcinomas (PCO). In pituitary αT3-1 cells, the expression level of hGnRHR was ten-fold higher than ovarian carcinoma OVCAR-3 cells. The possibility of the differential use of various cell-specific promoters in different cells was addressed by transiently transfecting cells with 3'-deletion clones of human GnRHR promoter. Sequential deletion of the 5'-flanking region of the gene revealed the use of two putative promoters, designated PR1 (-771 to -557) and PR2 (-1351 to -1022), and a negative control region (-1022 to -771), in the pituitary and ovarian cells. The same promoters appeared to be utilized for driving the basal promoter activities in both αT3-1 and OVCAR-3 cells, suggesting that there is no cell-specific promoter usage for the human GnRHR gene. Alternatively, the involvement of different regulatory protein factors was investigated using electrophoretic gel mobility shift assays. When end-labeled PR1 was used as a probe, two unique shifted complexes were identified in OVCAR-3 cells when compared to αT3-1 cells. One unique protein-DNA complex was observed in αT3-1 cells compared to OVCAR-3 cells when incubated with end-labeled PR2 as a probe. These DNA-protein complexes appeared to be specific, as they competed with excess amount of unlabelled competitor PR1 and PR2, respectively. In summary, it is unlikely that the use of a cell-specific promoter contributes to the different characteristics of ovarian GnRHR. Our study demonstrates that one mechanism by which cell-specific expression of human GnRHR is achieved is through the binding of distinct and/or combinations of cell-specific regulatory factors to various promoter elements in the 5'-flanking region of the gene. (C) 2000 Elsevier Science Ireland Ltd.en_HK
dc.languageengen_HK
dc.publisherElsevier Ireland Ltd. The Journal's web site is located at http://www.elsevier.com/locate/mceen_HK
dc.relation.ispartofMolecular and Cellular Endocrinologyen_HK
dc.rightsMolecular and Cellular Endocrinology. Copyright © Elsevier Ireland Ltd.en_HK
dc.subjectDifferential expressionen_HK
dc.subjectHuman GnRH receptoren_HK
dc.subjectOvaryen_HK
dc.subjectTranscription factorsen_HK
dc.titleDifferential expression of human gonadotropin-releasing hormone receptor gene in pituitary and ovarian cellsen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0303-7207&volume=162&spage=157&epage=166&date=2000&atitle=Differential+expression+of+human+gonadotropin-releasing+hormone+receptor+gene+in+pituitary+and+ovarian+cellsen_HK
dc.identifier.emailNgan, ESW: engan@hku.hken_HK
dc.identifier.emailChow, BKC: bkcc@hku.hken_HK
dc.identifier.authorityNgan, ESW=rp00422en_HK
dc.identifier.authorityChow, BKC=rp00681en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/S0303-7207(00)00196-9en_HK
dc.identifier.pmid10854709-
dc.identifier.scopuseid_2-s2.0-0343527941en_HK
dc.identifier.hkuros49325en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0343527941&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume162en_HK
dc.identifier.issue1-2en_HK
dc.identifier.spage157en_HK
dc.identifier.epage166en_HK
dc.identifier.isiWOS:000087642900018-
dc.publisher.placeIrelanden_HK
dc.identifier.scopusauthoridKang, SK=25623598700en_HK
dc.identifier.scopusauthoridCheng, KW=35081802000en_HK
dc.identifier.scopusauthoridNgan, ESW=22234827500en_HK
dc.identifier.scopusauthoridChow, BKC=7102826193en_HK
dc.identifier.scopusauthoridChoi, KC=7403949681en_HK
dc.identifier.scopusauthoridLeung, PCK=55419381000en_HK

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