Non-Surgical Gene Transfer to Rabbit Renal Glomeruli via Percutaneous Arterial Catheterization / 대한신장학회잡지

BACKGROUND: Transfer of foreign genes to the renal glomerular cells is an important step for the gene therapy of renal diseases in which the primary pathology is confined to the glomeruli. We developed a non-surgical method of gene transfer to rabbit renal glomeruli using percutaneous arterial catheterization without any laparatomy procedure. METHODS: The recombinant adenovirus type 5, containing a nuclear-targeted beta-galactosidase gene and driven by a cytomegalovirus promoter, was slowly infused into the unilateral renal artery via percutaneous arterial catheterization. The animals were sacrificed 3 days after virus infusion and lacZ staining was done on the fresh harvested tissue. RESULTS: Only the animals those received 6x10(12) particles/rabbit for 120 minutes show lacZ expression in 90.6+/-5% (n=3) of glomeruli. Mostly, it was the endothelial cells and mesangial cells those were positive for the stain. CONCLUSION: This non-surgical method for gene transduction of the renal glomeruli can be applied to human trials of glomerulus-directed gene therapy.

Involvement of mitogen-activated protein kinases and p21Waf1 in hydroxyurea-induced G1 arrest and senescence of McA-RH7777 rat hepatoma cell line

Hydroxyurea is commonly used to treat hematologic disorders and some type of solid tumors, but the mechanism for its therapeutic effect is not clearly known. In this study, we examined the effect of hydroxyurea on rat hepatoma McA-RH7777 cells, specifically, on the role of mitogen-activated protein (MAP) kinase signal transduction pathways and p21Waf1, p27Kip1 and p53. Rat hepatoma McA-RH7777 cells treated with hydroxyurea for 7 days, caused the inhibition of cell growth in a dose-dependent manner. But, this growth inhibition was not caused by necrosis or apoptosis but instead was associated with cell senescence-like change as evidenced by senescence associated-beta-galactosidase staining, and cells arrest at G1 phase of cell cycle. Phosphorylation of MAP kinases, such as ERK, JNK, and p38, was found to be decreased after treatment of cells with hydroxyurea. But, the expression of p21Waf1 was increased, while p27Kip1 and p53 were not detected in hydroxyurea treated rat hepatoma cells. Hydroxyurea treatment induced G1 arrest and a senescence-like changes in rat hepatoma McA-RH7777 cells may be the likely results of signal disruption of MAP kinases (ERK, JNK, and p38 MAP kinase) and p21Waf1 over-expression.