Glucocorticoid-regulated expression of exogenous human growth hormone gene in rats.
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.
Inazawa
T
T
Department of Advanced Medical Science, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan. inazawat@ims.u-tokyo.ac.jp
Tanabe
T
T
Yamada
H
H
Nakaoka
T
T
Hashimoto
Y
Y
Yamasaki
T
T
Kotaki
H
H
Tani
K
K
Asano
S
S
Yamashita
N
N
D000818:Animals; D002214:Capsules; D002460:Cell Line; D017690:Cell Transplantation; D003345:Corticosterone / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects*; D015316:Genetic Therapy / Q000379:methods; D005938:Glucocorticoids / Q000494:pharmacology*; D019382:Human Growth Hormone / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology; D008297:Male; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D015183:Restriction Mapping; D020079:Terminal Repeat Sequences / Q000235:genetics; D013997:Time Factors; D014162:Transfection
D002214:Capsules; D005938:Glucocorticoids; D019382:Human Growth Hormone; D003907:Dexamethasone; D003345:Corticosterone; D006854:Hydrocortisone