Effects of Autologous Serum and Platelet-Rich Plasma on Human Corneal Endothelial Cell Regeneration: A Comparative Study.

OBJECTIVES: To investigate the effects of autologous serum (AS) and platelet-rich plasma (PRP) on human corneal endothelial cell (HCEC) proliferation and apoptosis in comparison to Y-27632 as the commonly studied Rho-associated kinase (ROCK) inhibitor. METHODS: The human corneal endothelial primary cell line was used for this study. As the treatment groups, HCECs were incubated with AS, PRP, and Y-27632, whereas the control group received no treatment. Cell proliferation (measured by 5-bromo-2'-deoxyuridine [BrdU] incorporation) and apoptosis (based on the caspase-3 level) were compared between the control, Y-27632, AS, and PRP groups. RESULTS: In the Y-27632, AS, and PRP groups, the ratios of BrdU-incorporated cells were significantly higher (115±0.2%, 125±0.2%, 122±0.4% at 24 hr, and 138±2.4%, 160±0.2%, 142±0.2% at 48 hr, respectively) than in the control group (100±18.4% at 24 hr, 100±1.1% at 48 hr) ( P <0.05 for all). Furthermore, AS provided a higher HCEC proliferation ratio compared with the Y-27632 group at 24 and 48 hr ( P <0.05 for all). Caspase-3 was significantly lower in the AS group (60.3±3.3%) than in the control (100±2.3%), Y-27632 (101.9±5.2%), and PRP (101±6.8%) groups ( P <0.05 for all). CONCLUSIONS: The results of this study demonstrated for the first time that AS and PRP promoted HCEC proliferation and AS significantly decreased apoptosis in HCECs. A superior effect on HCEC proliferation was also observed with AS compared with Y-27632. Future "autologous" regenerative therapeutic options for corneal endothelial failure may involve the utilization of AS and PRP owing to their accessibility, simplicity in preparation, immunologic compatibility, and donor-free nature.

Investigating the Activity of Indole-2-on Derivative Src Kinase Inhibitors Against Chronic Myeloid Leukemia Cells.

BACKGROUND: Src family tyrosine kinases play a potential role in Bcr-Abl-induced leukemogenesis. Src kinase inhibitors are reported as selective inhibitors of chronic myeloid leukemia. OBJECTIVE: Since Src kinase inhibitors have an inhibitive effect on chronic myeloid leukemia, indole derivatives (C-1, C-2, C-3) previously found as potent inhibitors of Src kinase were tested against chronic myeloid leukemia in this study. METHODS: Cell viability of K562 and R/K562 cells, antiproliferative and antioxidant effects, and inhibition profiles of Bcr-Abl kinase of indole derivatives were determined compared to dasatinib and imatinib. RESULTS: The results showed that compounds affected cell proliferation and decreased the levels of Bcr/Abl. These results confirmed that the antileukemic activity of compounds was related to Bcr/Abl expression. Docking studies also presented that compounds are inhibitors of both Src and Abl kinases. Calculation of drug-like properties showed that compounds could be potential drug candidates. CONCLUSION: Among indole-2-on derivatives, previously identified as Src kinase inhibitors, C-2 has been discovered to be a strong anticancer drug that is active against susceptible and resistant K562 cell lines and induces apoptosis.

Verbascoside potentiates the effect of tyrosine kinase inhibitors on the induction of apoptosis and oxidative stress via the Abl-mediated MAPK signalling pathway in chronic myeloid leukaemia.

Verbascoside (Verb) may exhibit potential antitumour activities in leukaemia. The present study investigated the effect of Verb, in combination with imatinib (IM), dasatinib (Das), lipopolysaccharide (LPS) and TNF, on cell survival, Abl expression, apoptosis, oxidative stress and the MAPK pathway in chronic myeloid leukaemia (CML) cells. Cell viability was determined using the WST-8 assay in K562 and R-K562 cells treated with Verb and/or IM, Das, LPS and TNF. Apoptosis and DNA damage in CML cells was detected by caspase-3 and comet analysis. The protein levels of Abl (Phospho-Tyr412), and total/phosphorylated p38, JNK and ERK in CML cells were analysed using a Colorimetric Cell-Based Assay. Oxidative stress was examined using total antioxidant and oxidant status assays. Treatment with Verb and/or tyrosine kinase inhibitors (TKIs), LPS and TNF resulted in a significant decrease in the Tyr-412 phosphorylation of Abl in K562 and R-K562 cells. In addition, cotreatment with Verb and IM or Das additively induced apoptosis by activating caspase-3 levels in both cell lines. Activation of p38 and JNK can result in growth arrest and cell death, whereas ERK stimulation results in cell division and differentiation. The present study demonstrated that cotreatment with Verb and TKIs suppressed phosphorylated-ERK1/2, whereas the levels of phosphorylated-p-38 and phosphorylated-JNK were significantly elevated by Verb and/or IM, Das, LPS and TNF, thus suggesting that Abl and Src inhibition could be involved in the effects of Verb on MAPK signalling in R-K562 cells. Furthermore, Verb elevated reactive oxygen species levels additively with TKIs in both cell lines by increasing the oxidant capacity and decreasing the antioxidant capacity. In conclusion, anti-leukemic mechanisms of Verb may be mediated by Abl protein and regulation of its downstream p38-MAPK/JNK pathway, caspase-3 and oxidative stress in CML cells.