A human B-lymphoblastoid cell line produces prolactin
| Autor: | Birgit Gellersen, G E DiMattia, Heinz G. Bohnet, Henry G. Friesen |
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| Rok vydání: | 1988 |
| Předmět: |
Untranslated region
endocrine system medicine.medical_specialty endocrine system diseases Transcription Genetic Ribonuclease H Clone (cell biology) Radioimmunoassay Biology Cell Line Endocrinology Complementary DNA Internal medicine Endoribonucleases medicine Humans Northern blot RNA Messenger Southern blot Immunoassay Messenger RNA B-Lymphocytes Genes Immunoglobulin Lymphoblast Antibodies Monoclonal Nucleic Acid Hybridization DNA Molecular biology Prolactin Cell culture Immunoglobulin G Electrophoresis Polyacrylamide Gel Female Multiple Myeloma hormones hormone substitutes and hormone antagonists |
| Zdroj: | Endocrinology. 122(6) |
| ISSN: | 0013-7227 |
| Popis: | A variety of cell lines were examined by Northern blot hybridization for the expression of PRL or PRL-related mRNAs. We found that a human B-lymphoblast cell line transcribed a mRNA which hybridized to human PRL cDNA under high stringency conditions. The human lymphoblast cell line of interest is a variant subline of the IM-9 line that we have designated IM-9-P. The lymphoblast-derived PRL mRNA is approximately 150 bases longer than that produced by the human pituitary as determined by Northern blot analysis. IM-9-P PRL was immunoaffinity purified from conditioned medium and found to be identical in mol wt by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to human pituitary PRL. Moreover, IM-9-P PRL is biologically active in the rat Nb2 lymphoma mitogenic assay. Ribonuclease-H digestion of mRNA poly(A) tracts indicated that the size difference between pituitary and IM-9-P PRL transcripts was not due to an elongated poly(A) tail on the lymphoid PRL mRNA. Genomic Southern blot analysis showed no major rearrangements of the PRL gene in IM-9-P cells compared to the parent IM-9 line and human placenta DNA. Thus, it is highly likely that an elongation of the 5' and/or 3' untranslated regions of IM-9-P PRL mRNA account for the size difference with pituitary PRL mRNA. The PRL-producing IM-9-P line was cloned by limiting dilution, and a high PRL-producing clone IM-9-P3 and a non-PRL producer IM-9-P6 were isolated for further analysis. IM-9-P3 cells were found to secrete 40-50 ng PRL/10(6) cells.24 h regardless of cell density. The level of PRL mRNA also remained constant during exponential growth of IM-9-P3 cells. The existence of the PRL-producing IM-9-P3 clone and the IM-9-P6 clone which does not produce PRL as well as the IM-9 progenitor line provides a unique system with which to analyze the molecular mechanism of ectopic human PRL expression. |
| Databáze: | OpenAIRE |
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