The Use of Nanomedicine to Target Signaling by the PAK Kinases for Disease Treatment

P21-activated kinases (PAKs) are serine/threonine kinases involved in the regulation of cell survival, proliferation, inhibition of apoptosis, and the regulation of cell morphology. Some members of the PAK family are highly expressed in several types of cancer, and they have also been implicated in several other medical disorders. They are thus considered to be good targets for treatment of cancer and other diseases. Although there are several inhibitors of the PAKs, the utility of some of these inhibitors is reduced for several reasons, including limited metabolic stability. One way to overcome this problem is the use of nanoparticles, which have the potential to increase drug delivery. The overall goals of this review are to describe the roles for PAK kinases in cell signaling and disease, and to describe how the use of nanomedicine is a promising new method for administering PAK inhibitors for the purpose of disease treatment and research. We discuss some of the basic mechanisms behind nanomedicine technology, and we then describe how these techniques are being used to package and deliver PAK inhibitors.

High efficacy of tamoxifen-loaded L-lysine coated magnetic iron oxide nanoparticles in cell cycle arrest and anti-cancer activity for breast cancer therapy

Introduction: Due to the side effects of drugs, the development of nanoscale drug delivery systems has led to a significant improvement in medicinal therapies due to drug pharmacokinetics changes, decreased toxicity, and increased half-life of the drug. This study aimed to synthesize tamoxifen (TMX)-loaded L-lysine coated magnetic iron oxide nanoparticles as a nano-carrier to investigate its cytotoxic effects and anti-cancer properties against MCF-7 cancer cells. Methods: Magnetic Fe3O4 nanoparticles were synthesized and coated with L-lysine (F-Lys NPs). Then, TMX was loaded onto these NPs. The characteristics of synthesized nanoparticles (F-Lys-TMX NPs) were evaluated by X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS), differential scanning calorimetry (DSC), vibrating sample magnetometer (VSM), and thermogravimetric analysis (TGA). The drug release was analyzed at pH 5.8 and pH 7.4. The MCF-7 cells were exposed to F-Lys-TMX NPs, F-Lys NPs, and TMX for 24, 48, and 72 hours. To evaluate the cytotoxic potential of designed nanoparticles, MTT and apoptosis assays, real-time PCR, and cell cycle analysis was carried out. Results: The F-Lys-TMX NPs had spherical morphology with a size ranging from 9 to 30 nm. By increasing the nanoparticles concentration and treatment time, more cell proliferation inhibition and apoptosis induction were observed in F-Lys-TMX NPs-treated cells compared to the TMX. The expression levels of ERBB2, cyclin D1, and cyclin E genes were down-regulated and expression levels of the caspase-3 and caspase-9 genes were up-regulated. Studies on the drug release revealed a slow and controlled pH-dependent release of the nanoparticles. Cell cycle analysis indicated that F-Lys-TMX NPs could arrest the cells at the G0/G1 phase. Conclusion: The findings suggest that F-Lys-TMX NPs are more effective and have the potential for cell proliferation inhibition and apoptosis induction compared to the TMX. Hence, F-Lys-TMX NPs can be considered as an anti-cancer agent against MCF-7 breast cancer cells.

Cysteine Restriction in Murine L929 Fibroblasts as an Alternative Strategy to Methionine Restriction in Cancer Therapy

Methionine restriction (MetR) is an efficient method of amino acid restriction (AR) in cells and organisms that induces low energy metabolism (LEM) similar to caloric restriction (CR). The implementation of MetR as a therapy for cancer or other diseases is not simple since the elimination of a single amino acid in the diet is difficult. However, the in vivo turnover rate of cysteine is usually higher than the rate of intake through food. For this reason, every cell can enzymatically synthesize cysteine from methionine, which enables the use of specific enzymatic inhibitors. In this work, we analysed the potential of cysteine restriction (CysR) in the murine cell line L929. This study determined metabolic fingerprints using mass spectrometry (LC/MS). The profiles were compared with profiles created in an earlier work under MetR. The study was supplemented by proliferation studies using D-amino acid analogues and inhibitors of intracellular cysteine synthesis. CysR showed a proliferation inhibition potential comparable to that of MetR. However, the metabolic footprints differed significantly and showed that CysR does not induce classic LEM at the metabolic level. Nevertheless, CysR offers great potential as an alternative for decisive interventions in general and tumour metabolism at the metabolic level.

Antioxidant, Scavenging, Reducing, and Anti-Proliferative Activities of Selected Tropical Brown Seaweeds Confirm the Nutraceutical Potential of Spatoglossum asperum

Brown seaweeds have shown high potential of bioactivity and provide health benefits as an important functional food ingredient. Therefore, four abundantly growing tropical brown seaweeds—Iyengaria stellata, Spatoglossum asperum, Sargassum linearifolium, and Stoechospermum polypodioides—were collected from the Saurashtra Coast of the Arabian Sea. They were analyzed for metabolite profiling, biochemical activities (including total antioxidant, reducing, scavenging, and anti-proliferative characteristics), and total phenolic and flavonoid contents. A concentration-dependent antioxidant, reducing, and scavenging activities were observed for all four brown seaweeds. The S. asperum and I. stellata extracts showed maximum total antioxidant activity. S. asperum also showed high scavenging and reducing activities compared to other studied brown seaweeds. Further, S. asperum contained high total phenolic and flavonoid content compared to other brown seaweeds collected from the same coast. A multivariate correlation study confirmed a positive correlation between total phenolic and flavonoid contents, and biochemical activities (total antioxidant, scavenging and reducing) for all brown seaweeds. About 35% anti-proliferative activity was observed with S. asperum extract on Huh7 cells; in contrast S. polypodioide showed about 44% proliferation inhibition of Huh7 cells. Similarly, 26% proliferation inhibition of HeLa cells was observed with S. asperum extract. Overall, S. asperum possesses high total flavonoid and phenolic amounts, and showed potential antioxidant, scavenging and reducing characteristics. The study confirmed the nutraceutical potential of S. asperum and that it could be a promising functional food ingredient.

Study on the Antitumor Effect and Glycolysis of Andrographolide in Anaplastic Thyroid Carcinoma

Objective. To investigate the antitumor effect of andrographolide on the ATC cell lines 8505C and CAL62 and to explore the possible mechanism of the effect. Methods. CCK8 and colony formation assays were performed to detect proliferation. Cell migration was tested by scratch assay. Annexin V/PI staining was used to detect cell apoptosis and cell cycle. Glucose and lactic acid kits were carried out to evaluate the glycolysis level after andrographolide treatment. Western blot was used to detect the changes in the apoptosis-related proteins and glycolysis-related enzymes in both 8505C and CAL62 cells. Results. Treatment with 60 μM andrographolide had significant effects on 8505C and CAL62, including inhibition of proliferation, inhibition of migration, arrest of the cell cycle, promotion of apoptosis, and inhibition of glycolysis. Conclusion. Andrographolide has an antitumor effect and can significantly affect glycolysis in ATC cells.

Effects of different alcoholic beverages on human glioblastoma cell lines

Background: Anecdotal reports from neurosurgeons suggested that patients with glioblastoma who consumed a moderate amount of alcoholic beverages after glioblastoma surgery presented with improved vitality. Objective: This study aimed to investigate that if any evidence for these anecdotal reports can be reproduced experimentally. We also studied the effects of different alcoholic beverages on glioblastoma cells. Methods: GOS-3 glioblastoma cells and PC3 prostate carcinoma cells as control were incubated with beer, red wine, white wine, vodka, and whiskey at different concentrations. Membrane disruption by acute cytotoxicity and apoptosis induction was evaluated via Annexin-V-FITC flow cytometry and confocal laser scanning microscopy. Long-term effects on cell proliferation were studied by the XTT test. Results: There was no increase in membrane disruption even at physiologically high alcohol concentrations of 1 ‰. Cell proliferation was significantly inhibited by vodka and beer. Among the wines, the white wine caused slight proliferation inhibition in GOS-3 glioblastoma cells while inducing slightly enhanced proliferation in PC3 prostate cancer cells. After these results, more different brands of vodka and additional white and red wines from different grapes were used. While confirming the initial results, no additional differences between the different brands of vodka were observed. In the wine investigations, all the wines showed cell proliferation inhibition during long-term incubation of three different glioblastoma cell lines. Consistently, the inhibition from red wines was lower than the inhibition from white and rosé wines. Conclusion: In conclusion, alcoholic beverages at different concentrations used during the ingestion have both cytotoxic and antiproliferative effects on glioblastoma cells in vitro which could not be found in the controls with pure ethanol.