Isolation of genes differentially expressed at the downstream anastomosis of prosthetic arterial grafts with use of mRNA differential display
| Autor: | A D, Hamdan, L P, Aiello, W C, Quist, C K, Ozaki, M A, Contreras, M D, Phaneuf, C, Ruiz, G L, King, F W, LoGerfo |
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| Rok vydání: | 1995 |
| Předmět: |
DNA
Complementary Hyperplasia Base Sequence Transcription Genetic Anastomosis Surgical Retinoblastoma Gene Expression Blotting Northern Polymerase Chain Reaction Blood Vessel Prosthesis Interferon-gamma Carotid Arteries Dogs Genes Sequence Homology Nucleic Acid alpha 1-Antitrypsin Animals Aorta Abdominal RNA Messenger Cloning Molecular Creatine Kinase Polytetrafluoroethylene |
| Zdroj: | Journal of vascular surgery. 21(2) |
| ISSN: | 0741-5214 |
| Popis: | Downstream anastomotic intimal hyperplasia in prosthetic arterial grafts remains a major cause of delayed graft failure. The new method of messenger RNA (mRNA) differential display was used to screen numerous genes to gain insight into the molecular mechanisms of intimal hyperplasia.Fifty-centimeter-long 8 mm expanded polytetrafluoroethylene grafts were placed in four mongrel dogs from the carotid artery to the distal abdominal aorta. At 3 months the distal anastomoses and adjacent normal aortas were harvested; a portion was taken for histologic examination, and total RNA was isolated from the remainder. Differential mRNA display was used to identify candidate cDNA clones whose expression differed in anastomotic intimal hyperplasia as compared with adjacent unaffected aorta. The clones were sequenced, and national gene databases were searched. Northern blot analysis confirmed alteration of gene expression.Approximately 5000 mRNA species were screened, and 11 candidate clones were obtained. DNA sequence revealed homology of five clones to known gene sequences. Homologous genes included an interferon-gamma-induced human gene, (IGUP I-5111), alpha-1 protease inhibitor gene, human retinoblastoma susceptibility gene, and human creatine kinase gene (two clones). Northern blot analysis revealed altered gene expression in 4 of 11, nonregulation in 1 of 11, and undetectable signals in 6 of 11. Expression of the clone representing IGUP I-5111 in the segment of intimal hyperplasia was found to be decreased over threefold to only 31% +/- 16.4% SE of the level seen in normal aorta.The technique of mRNA differential display has identified differences in gene expression in an in vivo model of anastomotic intimal hyperplasia. Expression of RNA with homology to an interferon-gamma-induced human gene was consistently decreased within the hyperplastic region at the downstream polytetrafluoroethylene arterial anastomosis. |
| Databáze: | OpenAIRE |
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