Gene and protein profiling of the response of MA-10 Leydig tumor cells to human chorionic gonadotropin.
Activation of the steroidogenic machinery by peptide hormones involves a number of steps for transmitting signals from the plasma membrane to mitochondria in a spatially and temporally coordinated manner. Although key proteins mediating the hormonal signal have been identified, recent data suggest that the pathway might involve more complex protein-protein and protein-lipid interactions. Genomic and proteomic methods of analysis, namely the Affymetrix Murine Genome U74A v2 GeneChip and the BD PowerBlot Western Array, were used to identify human chorionic gonadotropin (hCG)-induced changes in mRNA and protein of MA-10 Leydig tumor cells that parallel the increase seen in progesterone synthesis. To analyze the massive amount of data that was generated, a comprehensive protein information matrix summarizing the features of each gene or protein, including its known properties, as well as annotations derived by homology-based functional inference, was developed. Of the genes examined by Affymetrix array, approximately 79 were differentially expressed and of gene products examined by PowerBlot, 9 were differentially expressed (above twofold). Changes in the expression of selected transcripts of interest were confirmed using real-time quantitative polymerase chain reaction and immunoblot analyses. Collectively, these results indicate that hormonal regulation of steroidogenesis is a complex phenomenon, involving proteins that participate in various known and novel pathways, which are implicated in transmitting signals from the plasma membrane to mitochondria and nucleus.
Li
Wenping
W
Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, Basic Sciences Bldg, 3900 Reservoir Rd, Washington, DC 20057, USA.
Amri
Hakima
H
Huang
Hongzhan
H
Wu
Cathy
C
Papadopoulos
Vassilios
V
D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D019295:Computational Biology; D020869:Gene Expression Profiling; D006801:Humans; D015151:Immunoblotting; D016247:Information Storage and Retrieval; D007984:Leydig Cell Tumor / Q000378:metabolism* / Q000473:pathology; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction / Q000379:methods; D011374:Progesterone / Q000096:biosynthesis; D040081:Protein Array Analysis; D013736:Testicular Neoplasms / Q000378:metabolism* / Q000473:pathology
D006063:Chorionic Gonadotropin; D011374:Progesterone