Uncoupling oxidative phosphorylation with 2,4-dinitrophenol promotes development of the adhesion phenotype.

Objective: To determine the effect of uncoupling oxidative phosphorylation with 2,4-dinitrophenol (DNP) on adhesion phenotype development.

Design: Prospective experimental study.

Setting: Academic medical center.

Growth suppression effects of recombinant adenovirus expressing human lactoferrin on cervical cancer in vitro and in vivo.

Human lactoferrin (hLF) is a multifunctional glycoprotein that can inhibit cancer growth. The molecular mechanism of hLF-induced tumor growth inhibition is incompletely understood. Moreover, the adenovirus vector-mediated hLF (Ad-hLF) gene therapy on cervical cancer has not been yet characterized.

Macrophages induce the adhesion phenotype in normal peritoneal fibroblasts.

Objective: To determine whether macrophages, exposed to hypoxia, stimulate primary cultures of fibroblasts to acquire the adhesion phenotype. The adhesion phenotype has been previously characterized, in part, by increased fibroblast expression of transforming growth factor (TGF) β1, vascular endothelial growth factor (VEGF), and type I collagen.

Design: Media collected from human macrophages cultured under hypoxic conditions (2% O(2)) were used to treat human peritoneal fibroblasts.

Dichloroacetate induces apoptosis of epithelial ovarian cancer cells through a mechanism involving modulation of oxidative stress.

Epithelial ovarian cancer (EOC) cells are under intrinsic oxidative stress, which alters metabolic activity and reduces apoptosis. Key oxidative stress enzymes, including myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS), are upregulated and colocalized in EOC cells. Oxidative stress is also regulated, in part, by superoxide dismutase (SOD) and hypoxia-inducible factor (HIF) 1a.

Reduction of hypoxia-induced angiogenesis in ovarian cancer cells by inhibition of HIF-1 alpha gene expression.

Purpose: The goal of this study was to investigate the effects of silencing HIF-1 alpha gene expression with specific small interfering RNA (siRNA) on VEGF production and angiogenesis in epithelial ovarian cancer (EOC) cells.

Methods: Two EOC cell lines, MDAH-2774 and SKOV-3, were cultured under normoxic (20% O(2)) and hypoxic (2% O(2)) conditions using standard techniques. After EOC cells were transfected with siRNA, HIF-1 alpha and VEGF mRNA levels were measured by real-time RT-PCR.

Exposure to polychlorinated biphenyls enhances lipid peroxidation in human normal peritoneal and adhesion fibroblasts: a potential role for myeloperoxidase.

Nitric oxide, superoxide, and lipid peroxidation (LPO) produced under oxidative stress may contribute to the development of postoperative adhesions. The objective of this study was to determine the effects of polychlorinated biphenyls (PCBs) on LPO, superoxide dismutase, myeloperoxidase (MPO), and nitrite/nitrate in human normal peritoneal and adhesion fibroblasts. PCB treatment reduced inducible nitric oxide synthase (iNOS) expression as well as levels of nitrite/nitrate in both cell lines.

Myeloperoxidase serves as a redox switch that regulates apoptosis in epithelial ovarian cancer.

Objectives: Resistance to apoptosis is a key feature of cancer cells and is believed to be regulated by nitrosonium ion (NO(+))-induced S-nitrosylation of key enzymes. Nitric oxide (NO), produced by inducible nitric oxide synthase (iNOS), is utilized by MPO to generated NO(+). We sought to investigate the expression of myeloperoxidase (MPO) and iNOS in epithelial ovarian cancer (EOC) and determine their effect on S-nitrosylation of caspase-3 and its activity as well as apoptosis.

S-nitrosylation of caspase-3 is the mechanism by which adhesion fibroblasts manifest lower apoptosis.

We have previously found that adhesion fibroblasts exhibit lower apoptosis and higher protein nitration as compared with normal peritoneal fibroblasts. In this study, we sought to determine whether the decreased apoptosis observed in adhesion fibroblasts is caused by lower caspase-3 activity due to an increase in caspase-3 S-nitrosylation. For this study, we have utilized primary cultures of fibroblasts obtained from normal peritoneum and adhesion tissues of the same patient(s).

The effect of estradiol on the expression of estrogen, progesterone, androgen, and prolactin receptors in human peritoneal fibroblasts.

Purpose: To assess the ability of fibroblasts isolated from normal peritoneum and adhesion tissues to express various hormone receptors when cultured with exogenous estradiol.

Methods: Primary cultures of fibroblasts from normal human peritoneum and adhesion tissue were treated with zero (control), 10(-10), 10(-8), and 10(-6) M concentrations of 17beta-estradiol. We performed real time reverse transcriptase polymerase chain reaction to determine mRNA levels of estradiol-alpha receptor (ER-alpha) and estradiol-beta receptor (ER-beta), progesterone receptor (P-R), androgen receptor (A-R), and prolactin receptor (PRL-R) in the two types of fibroblast cultures.

Hypoxia-generated superoxide induces the development of the adhesion phenotype.

Adhesion fibroblasts exhibit higher TGF-beta1 and type I collagen expression as compared to normal peritoneal fibroblasts. Furthermore, exposure of normal peritoneal fibroblasts to hypoxia results in an irreversible increase in TGF-beta1 and type I collagen. We postulated that the mechanism by which hypoxia induced the adhesion phenotype is through the production of superoxide either directly or through the formation of peroxynitrite.

High-level expression of recombinant human nerve growth factor beta in milk of nontransgenic rabbits.

The technology for the large-scale production of therapeutic recombinant proteins remains a challenge in the biopharmaceutical industry. In this study, we reported a nontransgenic approach to producing a large quantity of human nerve growth factor beta (hNGF-beta) in rabbit milk by employing a recombinant adenoviral expression system. After directly instilling hNGF-beta recombinant adenoviruses into rabbit mammary glands, a polypeptide with a molecular weight of 13.

Nitric oxide synthase isoforms expression in fibroblasts isolated from human normal peritoneum and adhesion tissues.

Objective: To determine the expression of nitric oxide synthases (NOSs) and their modulation by hypoxia in human peritoneal (NF) and adhesion fibroblasts (ADF).

Design: Prospective experimental study.

Setting: University medical center.

Hypoxia regulates iNOS expression in human normal peritoneal and adhesion fibroblasts through nuclear factor kappa B activation mechanism.

Objective: To determine the mechanism by which hypoxia increases expression of iNOS in human normal peritoneal and adhesion fibroblasts.

Design: Prospective experimental study.

Setting: University medical center.