Detection of gonadotropin-releasing hormone receptor and its mRNA in primary human epithelial ovarian cancers
Journal
International journal of gynecological cancer : official journal of the International Gynecological Cancer Society
Journal Volume
14
Journal Issue
3
Date Issued
2004
Author(s)
Abstract
The hypothalamic neuropeptide gonadotropin-releasing hormone (GnRH) serves a key role in regulating mammalian reproductive function. An extrapituitary role for GnRH in the normal and malignant reproductive tissues has been postulated. The purpose of our study is to demonstrate the presence and levels of GnRH receptor (RGnRH) protein and its mRNA in normal and malignant tissues of ovary. Normal human ovarian tissues (n = 13), as well as epithelial ovarian cancer specimens from stages I-IV (n = 39), were obtained from appropriate patients at operation room. Monoclonal antibodies against RGnRH were used for immunohistochemical evaluation of paraffin-embedded ovarian tissue sections by methods of streptavidin-biotin immunostaining. The molecular size and levels of RGnRH were determined by enhanced chemiluminescence-Western blot assay. The amount of RGnRH mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR). The rate of positive immunostaining in ovarian cancers was 53.8% (21/39). The rate of positive staining in the late stage (stages III and IV) was significantly higher than that in the early stage (stages I and II). A single band of molecular weight of about 60 kDa was detected from protein extracts of ovarian cancer as well as from normal ovary. The mean values of fold increase of signal intensities of 60 kDa detected by Western blots in stages I-IV ovarian cancers were 2.39, 2.42, 2.78, and 3.62, respectively, as compared with normal ovarian tissues. The overall positive rate of Western blot analysis for ovarian cancers was 59% (23/39). The mean values of signal intensity of RT-PCR products of RGnRH mRNA in stages I-IV were 2.24, 2.58, 3.10, and 3.20, respectively. The positive rate of overexpression of RGnRH mRNA in ovarian cancer was 70% (21/30). The differences of mean values of signal intensities of Western blot staining (2.41 versus 2.85) as well as RT-PCR products (2.40 versus 3.11) between the early stage and the late stage of ovarian cancers were statistically nonsignificant. Mechanism of autocrine regulation of tumor growth in human epithelial ovarian cancer can be explained by the coexistence of GnRH, RGnRH, and its mRNA, according to our own and other studies. The level of RGnRH expressed by ovarian cancer might be used for targeting chemotherapeutic agents to those patients who harbor RGnRH-positive tumors.
Subjects
Immunohistochemistry | Ovarian cancer | RGnRH | RGnRH mRNA | RT-PCR | Western blot
SDGs
Other Subjects
biotin; gonadorelin receptor; messenger RNA; monoclonal antibody; neuropeptide; streptavidin; article; autocrine effect; cancer patient; chemoluminescence; epithelium tumor; female; human; human tissue; immunohistochemistry; ovary cancer; priority journal; protein expression; protein isolation; reverse transcription polymerase chain reaction; tissue section; tumor growth; Western blotting; Blotting, Western; Case-Control Studies; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Neoplasm Staging; Neoplasms, Glandular and Epithelial; Ovarian Neoplasms; Receptors, LHRH; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger
Type
journal article