Glucocorticoid-regulated expression of exogenous human growth hormone gene in rats

Mol Ther. 2001 Sep;4(3):267-72. doi: 10.1006/mthe.2001.0438.

Abstract

The aim of this study was to control in vitro and in vivo expression of the growth hormone (GH) gene using a glucocorticoid-sensitive promoter, the mouse mammary tumor virus long terminal repeat (MMTV LTR). We inserted the cDNA encoding the 20-kDa form of human GH (20K-GH) downstream of the MMTV LTR of plasmid pMSG, and used lipofection to transfer it to 3Y1 cells together with plasmid pMX, which contains a puromycin-resistant element. The secretion of GH from the selected transformants was dose-dependently augmented by the application of hydrocortisone, corticosterone, or dexamethasone, among which dexamethasone was the most potent. Analysis of the time course showed that 20K-GH secretion began to increase within 2 hours after the addition of glucocorticoid and reached a maximal level of about threefold over the unstimulated control at 3 hours; secretion then gradually declined and returned to near basal levels at 19 hours. Repeated glucocorticoid application led to repeated increases in GH secretion. When GH-producing cells were microcapsulated and transplanted into the abdominal cavities of rats, 20K-GH was detected in the plasma under control conditions and increased about 3.3-fold after administration of dexamethasone. We suggest that GH expression driven by the MMTV LTR promoter may be under the control of an endogenous glucocorticoid in vivo.

MeSH terms

  • Animals
  • Capsules
  • Cell Line
  • Cell Transplantation
  • Corticosterone / pharmacology
  • Dexamethasone / pharmacology
  • Dose-Response Relationship, Drug
  • Gene Expression Regulation / drug effects*
  • Genetic Therapy / methods
  • Glucocorticoids / pharmacology*
  • Human Growth Hormone / genetics*
  • Human Growth Hormone / metabolism*
  • Humans
  • Hydrocortisone / pharmacology
  • Male
  • Mammary Tumor Virus, Mouse / genetics
  • Promoter Regions, Genetic / genetics
  • Rats
  • Restriction Mapping
  • Terminal Repeat Sequences / genetics
  • Time Factors
  • Transfection

Substances

  • Capsules
  • Glucocorticoids
  • Human Growth Hormone
  • Dexamethasone
  • Corticosterone
  • Hydrocortisone