Gene Therapy Using Hepatocyte Growth Factor Expressing Adenovirus Improves Skin Flap Survival in a Rat Model

Hepatocyte growth factor (HGF) is a potent angiogenic factor that can stimulate the production of blood vessels in ischemic tissue. We investigated whether gene therapy using HGF-expressing adenovirus could enhance skin flap survival. Sprague-Dawley rats were randomly divided into three groups. Rats were subdermally injected with HGF-expressing adenovirus (HGF virus group), recombinant HGF (rhHGF group), or phosphate buffered saline (PBS group) 2 days before and immediately after 3 x 9 cm caudal flap elevation. The survival area of the skin flap, the ratio of blood flow, CD31-positive vessels and, VEGF expression were examined. Skin flap viability was significantly increased in the HGF virus group compared to the rhHGF and PBS groups (71.4% +/- 5.9%, 63.8%+/- 6.4%, and 39.2% +/- 13.0%, respectively) (P = 0.025). Furthermore, the blood flow ratio was significantly increased in the HGF virus group. In the HGF virus group, the number of CD31-positive vessels and vascular endothelial growth factor (VEGF) expression were significantly increased. Gene therapy using HGF-expressing adenovirus increase VEGF expression, the number of viable capillaries, and blood flow to the flap, thereby improving skin flap survival.

Evaluation of immunohistochemical staining for hepatocyte growth factor and c-met in endometrial adenocarcinoma

Endometrial carcinoma is a primary malignant epithelial tumor, usually with glandular differentiation, arising in the endometrium with the potential to invade and metastasize. Hepatocyte growth factor [HGF] and its receptor c- Met have been implicated in uterine development, pregnancy, and endometrial disorders, such as endometriosis and carcinoma. The goal of this study was to evaluate immunohistochemical [IHC] staining patterns of HGF and c-Met in endometrial adenocarcinoma and to correlate staining with the biological behavior and outcome of endometrial adenocarcinoma. A retrospective study included 45 cases, who underwent total abdominal hysterectomy and bilateral salpingo-oopherectomy, between 2005-2010 samples, were taken from Teaching Laboratories at Baghdad Teaching Hospital/Medical City. Thirty cases were diagnosed as adenocarcinoma of the endometrium, an additional 15 patients diagnosed as to have uterine leiomyoma [fibroid] and who had normal endometrium were taken as a control group. The patient's age, tumor grading, depth of myometrial invasion, presence of pelvic and paraaortic LN metstases, vascular invasion, and the stage of the disease were noted. The specimens were already fixed in10% formalin, and paraffin embedded. Three sections [4 micron in thickness] were cut from each paraffin block. One section stained with Haematoxylin and Eosin [H and E] stain, and the other two stained with HGF and c-Met immunostaining antibodies using positively charged slides. The mean age of the patients with endometrial adenocarcinoma was 58.5years. There was no statistically significant difference between HGF/c-Met expression and age, tumor grade, stage, myometrial invasion and vascular invasion. There was a statistical significant correlation between HGF and c-Met scores with cases showing no pelvic lymph nodes metastases. There is a linear increase in HGF and c-Met expression in both the diseased and control group. A statistical significant correlation was found in HGF and c-Met scores between the diseased group and the control group. HGF and c-Met staining was significantly different between control group and diseased group. HGF and c-Met Showed Linear increase Expression in both diseased and control group. The c-Met is the high-affinity receptor for hepatocyte growth factor.

Role of HGF/c-Met in Serum-Starved ARPE-19 Cells

PURPOSE: Hepatocyte growth factor (HGF) and its receptor (HGFR/c-Met) regulate motility, mitogenesis, and morphogenesis in a cell type-dependent fashion. We report the role of HGF and c-Met on stress-induced ARPE-19 human retinal pigment epithelial (RPE) cells in this study. METHODS: The cells were cultured either with or without serum. Southern and Western blot analyses were done to determine the expression patterns of HGF/c-Met in serum-starved ARPE-19 cells. The cell proliferation pattern in serum-starved condition was analyzed using MTS assay. Inhibition level of cell proliferation was analyzed using a neutralizing monoclonal antibody against c-Met (2 microgram/ml). RESULTS: Abnormal cell proliferation and scattering of ARPE-19 cells was observed under serum starvation. HGF/c-Met were expressed in serum-starved ARPE-19 cells. ARPE-19 cell proliferation was also enhanced with recombinant HGF treatment. Neutralization against c-Met inhibited the proliferation of serum-deprived ARPE-19 by 64.5% (n=9, S.D. 5.5%). Serum starvation appears to induce epithelial-mesenchymal transition of ARPE-19 cells, resulting in scatter, and the expression of alpha-smooth muscle actin (alpha-SMA), a marker for fibrosis. CONCLUSIONS: In conclusion, c-Met induced under non-physiologic conditions has significant effects on the activation of RPE cells.