Transduction of exogenous constitutively activated Stat3 into dispersed islets induces proliferation of rat pancreatic beta-cells.
In vitro proliferation of functional islet mass/beta-cells by transduction of cell cycle regulatory genes has not been well documented due to the lack of either an effective method for gene transduction to islets or effective genes for beta-cell proliferation. Signal transducers and activators of transcription- 3 (Stat3) are among the most important molecules for cell proliferation and for cell antiapoptosis. In this study, adenovirus-mediated gene transduction was performed on dispersed islet cells, and reaggregated islet mass functions, such as capabilities for reaggregation, proliferation, and glucose- stimulated insulin secretion (GSIS), were evaluated. The constitutively activated form of Stat3 (Stat3-C) was effectively transduced to most of the islet cells, even at a low density of adenovirus vector. There was no difference in the spontaneous reaggregation capability between Stat3-C transduced and control islet cells. BrdU incorporation in Stat3-C-transduced islet cells was significantly higher (2.8-fold) than that in the control cells, and it was significantly elevated (4-fold) by the addition of HGF in culture media. GSIS in Stat3-C-transduced islet cells was preserved to the level in controls by 5 days in culture. The results indicated that gene transduction into islet cells could be enhanced by first dispersing the cells. Stat3-C induced cell proliferation of beta-cells without loss of insulin secretion activity at the glucose challenge, and HGF enhanced the beta-cell proliferative activity of Stat3-C.
Tsukiyama
Shusaku
S
First Department of Surgery, Hokkaido University Graduate School of Medicine, Kita-ku, Sapporo, Japan.
Matsushita
Michiaki
M
Matsumoto
Shuichiro
S
Morita
Tsunehiko
T
Kobayashi
Satoru
S
Tamura
Hitosi
H
Kamachi
Hirohumi
H
Ozaki
Michitaka
M
Todo
Satoru
S
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D004357:Drug Synergism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005947:Glucose / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D050796:STAT3 Transcription Factor / Q000235:genetics* / Q000378:metabolism*; D014161:Transduction, Genetic
D006133:Growth Substances; D050796:STAT3 Transcription Factor; C494088:Stat3 protein, rat; D004815:Epidermal Growth Factor; D017228:Hepatocyte Growth Factor; D005947:Glucose