The full agonistic effect of recombinant 20 kDa human growth hormone (hGH) on CHO cells stably transfected with hGH receptor cDNA

Wada, Mayuko; Ikeda, Miwa; Takahashi, Yuka; Asada, Nobuhiko; Chang, Kyu‐Tae; Takahashi, Michio; Honjo, Masaru

doi:10.1016/s0303-7207(97)00151-2

Cited by 36 publications

(23 citation statements)

References 34 publications

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“…stimulating [3H]thymidine incorporation, Al-P activity and osteocalcin production at 0.5-5.0 nM, namely at the physiological concentration of the hormones in the human blood [2]. These in vitro findings are consistent with the recent observation that 22K-and 20K-hGH have identical binding affinities (0.41 nM) and exert full agonistic effects on CHO cells transf ected with hGH receptor cDNA [16,17]. Furthermore, the present in vitro findings are in keeping with the in vivo data showing that 20K hGH has the same weight gain activity as 22K hGH in hypophysectomized rats [16].…”

Section: Effects Of 22k-and 20k-hgh On Osteocalcin Productionsupporting

confidence: 88%

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Osteo-Anabolic Effects of Human Growth Hormone with 22K- and 20K Daltons on Human Osteoblast-Like Cells.

et al. 1999

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Section: Effects Of 22k-and 20k-hgh On Osteocalcin Productionsupporting

confidence: 88%

“…[12][13][14][15]. Recently, recombinant 20K-hGH with a natural sequence was synthesized and it was found that the recombinant 20K-and 22K hGH increases body weight in rats equipotently [16] and that both hGHs have the same affinity constant for hGH receptors transiently expressed on CHO cells [17].…”

mentioning

confidence: 99%

Osteo-Anabolic Effects of Human Growth Hormone with 22K- and 20K Daltons on Human Osteoblast-Like Cells.

et al. 1999

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“…1a). The expression of functional hGHR by transfecting this sequence into CHO cells has been confirmed elsewhere [22,23]. The NheI-EcoRI and NheI-Xho-I fragments of this construct were used as probes for Southern and Northern blot analyses.…”

Section: Preparations Of Transgene Constructionmentioning

confidence: 90%

Production of Transgenic Rats and Mice by the Testis-Mediated Gene Transfer

Chang

Ikeda

Hayashi

et al. 1999

Journal of Reproduction and Development

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Abstract. Recent reports have shown that sperm cells incubated with foreign DNA in vitro are able to transfer the DNA into eggs at fertilization. The present study examined if an injection of DNA into the testis in vivo could generate transgenic animals via sperm ejaculated. We prepared 3 gene constructs; human growth hormone (hGH), hGH receptor (hGHR) and mouse leptin (mOB) genes fused to the promoter regions of the mouse genes for whey acidic protein (WAP), metallothionein-I (MT) and mouse mammary tumor virus (MMTV), respectively. Each gene construct mixed with cationic liposome was injected into bilateral testes of male rats or mice, and the males mated with females 3 or 4 days later. A female rat mated with a male treated with MT/hGHR gene gave birth to 17 pups, 3 of which were found to carry the transgene. The expression of hGHR mRNA was demonstrated in the liver, kidney, muscle and brain after treating with zinc in drinking water. At present, the transmission of the exogenous gene to the descendants was confirmed up to the F4 generation. Three female rats mated with 3 different males injected with MMTV/mOB fusion gene produced 10, 14 and 12 pups, respectively. The genome of 2 out of these 36 pups harbored MMTVmOB gene, though expression of mOB mRNA was not detected. Two female mice were mated with 2 male mice injected with WAP/hGH gene and produced 15 and 16 pups, 3 of which incorporated the gene. The expression of hGH mRNA in the mammary gland in these mice was confirmed. These results indicate that exogenous DNA injected into the testis as a liposome-complex can be transferred into eggs via sperm and expressed in the postpartum progeny.

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“…The cells were reseeded into 25 cm 2 culture flasks (Nunc) and precultured for 24 h in a humidified incubator at 37~ and 5% CO2/95% air, before E2 (100 ng/ml) and/or hPRL (1 or 100 ng/ml), 22k hGH (1 or 100 ng/ml)or 20K hGH (1 or 100 ng/ml) were added. Northern Blot Analysis of Cyclin D1 mRNA---RNA was exracted by an acidic guanidinium isothiacyanate-phenol-chloroform (AGPC) procedure from T-47D cells which were harvested and pooled from triplicate flasks, and Northern blot analysis was performed with 20 pg of total RNA per lane as described previously (14). The membranes were hybridized at 40~ overnight with cyclin DI cDNA probe (generously provided by Dr. D. Beach, Howard Hughes Medical 9 32 Institute, Cold Spring Habor, NY) being labeled with [ct-P]dCTP (specific activity 111 Tbq/nmol, ICN Biochemicals, CA) to a final specific activity of 8 x 10 9 cpm/t*g of DNA by using a DNA labeling kit (Nippon Gene Co., Toyama, Japan).…”

Section: Introductionmentioning

confidence: 99%

Cooperative and differential effects of estrogen, prolactin, 22K and 20K human growth hormones on cyclin D1/PRAD1 gene expression in T‐47D human breast cancer cells

et al. 1998

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SUMMARYThe cyclin D1/PRAD1 gene is correlated with carcinogenesis of human breast cancer. In this study, we have analyzed effects of breast cancer-related hormones on the cyclin D1 gene expression in T-47D human breast cancer cells. Estradiol (E2) and human prolactin (hPRL) equally enhanced the cyclin D1 gene expression in the cells, and 22 and 20 kDa human growth hormones (22K and 20K hGHs) showed less stimulatory effects. In the presence of F~.however, hPRL or 22K hGH showed additive stimulations of the cyclin D1 gene expression to that by ~ alone, while 20K hGH did not show any additive stimulation of the gene expression. The results suggest that the signal pathways through estrogen and hPRL receptors are important for cyclin D1 gene expression in breast cancer cells, and that 20K hGH has little effect on the cyclin D1 gene expression in these cells because of its lower affinity to PRL receptor.

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The full agonistic effect of recombinant 20 kDa human growth hormone (hGH) on CHO cells stably transfected with hGH receptor cDNA

Cited by 36 publications

References 34 publications

Osteo-Anabolic Effects of Human Growth Hormone with 22K- and 20K Daltons on Human Osteoblast-Like Cells.

Osteo-Anabolic Effects of Human Growth Hormone with 22K- and 20K Daltons on Human Osteoblast-Like Cells.

Production of Transgenic Rats and Mice by the Testis-Mediated Gene Transfer

Cooperative and differential effects of estrogen, prolactin, 22K and 20K human growth hormones on cyclin D1/PRAD1 gene expression in T‐47D human breast cancer cells

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