RHOA and mDia1 Promotes Apoptosis of Breast Cancer Cells Via a High Dose of Doxorubicin Treatment.

BACKGROUND: Transforming RhoA proteins (RHOA) and their downstream Diaphanous homolog 1 proteins (DIAPH1) or mDia1 participate in the regulation of actin cytoskeleton which plays critical role in cells, i.e., morphologic changes and apoptosis. METHODOLOGY: To determine the cell viability the real time cell analysis (RTCA) and flow cytometry were used. To perform proteomic analysis, the label-free quantitative method and post-translation modification by the nano-HPLC and ESI-MS ion trap mass analyser were used. RESULTS: The results of the cell viability showed an increase of dead cells (around 30 %) in MCF-7/DOX-1 (i.e., 1µM of doxorubicin was added to MCF-7/WT breast cancer cell line) compared to MCF-7/WT (control) after 24 h doxorubicin (DOX) treatment. The signalling pathway of the Regulation of actin cytoskeleton (p<0.0026) was determined, where RHOA and mDia1 proteins were up-regulated. Also, post-translational modification analysis of these proteins in MCF-7/DOX-1 cells revealed dysregulation of the actin cytoskeleton, specifically the collapse of actin stress fibbers due to phosphorylation of RHOA at serine 188 and mDia1 at serine 22, resulting in their deactivation and cell apoptosis. CONCLUSION: These results pointed to an assumed role of DOX to dysregulation of actin cytoskeleton and cell death.

Effect of equal daily doses achieved by different power densities of low-level laser therapy at 635 nm on open skin wound healing in normal and diabetic rats.

BACKGROUND AND OBJECTIVE: Despite the fact that the molecular mechanism of low-level laser therapy (LLLT) is not yet known, the exploitation of phototherapy in clinical medicine and surgery is of great interest. The present study investigates the effects of LLLT on open skin wound healing in normal and diabetic rats. MATERIALS AND METHODS: Four round full-thickness skin wounds on dorsum were performed in male adult nondiabetic (n = 24) and diabetic (n = 24) Sprague-Dawley rats. AlGaInP (635 nm, wavelength; 5 J/cm(2), daily dose) was used to deliver power densities of 1, 5, and 15 mW/cm(2) three times daily until euthanasia. RESULTS: PMNL infiltration was lower in the irradiated groups (15 mW/cm(2)). The synthesis and organisation of collagen fibres were consecutively enhanced in the 5 mW/cm(2) and 15 mW/cm(2) groups compared to the others in nondiabetic rats. In the diabetic group the only significant difference was recorded in the ratio PMNL/Ma at 15 mW/cm(2). A significant difference in the number of newly formed capillaries in the irradiated group (5, 15 mW/cm(2)) was recorded on day six after injury compared to the control group. CONCLUSION: LLLT confers a protective effect against excessive inflammatory tissue response; it stimulates neovascularization and the early formation of collagen fibres.

Impact of different pancreatic microenvironments on improvement in hyperglycemia and insulin deficiency in diabetic rats after transplantation of allogeneic mesenchymal stromal cells.

BACKGROUND: Mesenchymal stromal cells (MSCs) in the pancreatic microenvironment can improve diabetes mellitus (DM). The aim of the present study was to determine whether different pancreatic microenvironments influence the improvement of hyperglycemia and insulin deficiency. METHODS: MSCs isolated from rat bone marrow were transplanted directly into different pancreatic microenvironments in male DM rats. DM was induced in the rats by streptozotocin injection. The rats were divided into 5 groups: normal control rats, DM control rats, and 3 experimental groups (DM rats plus MSCs injected into the head of the pancreas, the tail of the pancreas, or the whole pancreas). The body weight and blood glucose of the rats were monitored during the experiment after transplantation of the MSCs. Histopathologic and immunohistochemical analyses were used to detect the presence and number of islets and insulin production in the pancreatic tissue of the rats after MSC transplantation. RESULTS: At 28 days after MSC transplantation, we observed a statistically significant decrease in the blood glucose level and an increase in weight in DM rats compared with DM control rats (P < 0.0001 and P < 0.03, respectively). A comparison of each of the DM rat groups treated with MSCs showed no significant differences in the blood glucose levels or body weight. CONCLUSION: Our results suggest that transplantation of MSCs could improve DM in the pancreatic microenvironment in an animal model with streptozotocin-induced DM. The different pancreatic areas into which the MSCs were implanted had no significant influence on the improvement in hyperglycemia and insulin deficiency.

Effect of equal daily doses achieved by different power densities of low-level laser therapy at 635 nm on open skin wound healing in normal and corticosteroid-treated rats.

Optimal parameters of low-level laser therapy (LLLT) for wound healing are still discussed. Hence, our study was aimed to compare effects of different power densities of LLLT at 635 nm in rats. Four, round, full-thickness, skin wounds were made on the backs of 48 rats that were divided into two groups (non-steroid laser-treated and steroid laser-treated). Three wounds were stimulated daily with a diode laser (daily dose 5 J/cm(2)) each with different power density (1 mW/cm(2), 5 mW/cm(2), and 15 mW/cm(2)), whereas the fourth wound served as a control. Two days, 6 days, and 14 days after surgery, eight animals from each group were killed and samples were removed for histological evaluation. In the non-steroid laser-treated rats, significant acceleration of epithelization and collagen synthesis 2 days and 6 days after surgery was observed in stimulated wounds. In steroid laser-treated rats, 2 days and 14 days after surgery, a decreased leucocyte/macrophage ratio and a reduction in the area of granulation tissue were recorded, respectively. In conclusion, LLLT, by the method we used, improved wound healing in the non-steroid laser-treated rats, but it was useless after corticosteroid treatment.