Inhibition of bone morphogenetic proteins signaling suppresses metastasis melanoma: a proteomics approach.

BACKGROUND: Bone morphogenetic proteins (BMPs) are members of the transforming growth factor-ß superfamily, known to promote the tumor invasion and metastasis. There are continual progresses in understanding the role of BMP signaling pathways in carcinogenesis. However, the biological significance of BMPs in human melanoma has received very little attention. The study aimed to explore the effect of BMP inhibition on melanoma treated with LDN193189 (BMP inhibitor) using a quantitative proteomics approach in a melanoma xenograft model. MATERIALS AND METHODS: Melanoma tumor was induced in C57BL6 mice and treated intraperitoneally with LDN193189 for ten consecutive days. Post-treatment, tumors were collected, and comparative proteomics was performed using a high-resolution Orbitrap Fusion Tribrid mass spectrometer. RESULTS: Treatment of melanoma with LDN193189 at 3 mg/kg body weight twice daily showed a significant decrease in the growth rate of the tumor compared to the other doses tested. Quantitative proteomic profiling identified 3231 proteins. Bioinformatics analysis of the 131 differentially expressed proteins selected by their relative abundance revealed that LDN193189 induces alterations in the cellular and metabolic process and the proteins that are involved in protein binding and catalytic activity in melanoma. CONCLUSIONS: Down-regulation of metallothionein (MT) 1 and MT2, emerging proteins for their role in tumor formation, progression, and drug resistance and transcription factor EB that plays a crucial role in the regulation of basic cellular processes, such as lysosomal biogenesis and autophagy, were identified upon inhibition of the BMP pathway in melanoma, suggesting their roles in melanoma growth. Understanding the role of these proteins will provide new directions for treating cancer.

Physiological and biochemical responses of Amaranthus cruentus to polycyclic aromatic hydrocarbon pollution caused by thermal power units.

Pollution due to release of polycyclic aromatic hydrocarbons from thermal power plants is a major global issue as the same is highly toxic and carcinogenic. The current research aims to investigate the responses of a dietary plant Amaranthus cruentus towards PAH pollution. For the said purpose, the plant was collected from agricultural land in close vicinity to thermal power units and the effects of PAH pollution on its chlorophyll and various nutraceutical content was evaluated. Oxidative stress biomarkers and antioxidant defense enzymes status and PAH accumulation was quantified as well. Real-time evidence of cell death, depletion of nutraceutical resources, and stomata configuration was generated through various histochemical studies and SEM analysis. Results indicated significant decline of chlorophyll a to the extent of 77% when compared to control. Oxidative stress markers, namely, superoxide radical, H2O2, and hydroxyl radical in pollution exposed plants were 12.7, 2.2, and 2.4 times respectively higher over the control which eventually resulted in 35% more cell death for the pollution exposed group. Total phenolics and flavonoids showed a decline of 57.6% and 41.3% respectively in the group exposed to PAH pollution. Similar decreasing trend was also observed for ascorbic acid, α-tocopherol, ß-carotene, total proteins, and carbohydrate contents as well. PAH-induced stress also resulted in complete imbalance in the redox homeostasis of the plant which was evident from increase in super oxide dismutase, catalase, and peroxidase antioxidant enzymes by more than 2-fold when compared to control. PAH accumulation in sample group was 10-20 times more when compared to control. Proteomic analysis also indicated upregulation of some proteins related to stress situation. Results are evident of the fact that severe depletion of nutraceutical resources of dietary plants can take place if subjected to oxidative stress arising from PAH pollution.

A mycobacterial phosphoribosyltransferase promotes bacillary survival by inhibiting oxidative stress and autophagy pathways in macrophages and zebrafish.

Mycobacterium tuberculosis employs various strategies to modulate host immune responses to facilitate its persistence in macrophages. The M. tuberculosis cell wall contains numerous glycoproteins with unknown roles in pathogenesis. Here, by using Concanavalin A and LC-MS analysis, we identified a novel mannosylated glycoprotein phosphoribosyltransferase, encoded by Rv3242c from M. tuberculosis cell walls. Homology modeling, bioinformatic analyses, and an assay of phosphoribosyltransferase activity in Mycobacterium smegmatis expressing recombinant Rv3242c (MsmRv3242c) confirmed the mass spectrometry data. Using Mycobacterium marinum-zebrafish and the surrogate MsmRv3242c infection models, we proved that phosphoribosyltransferase is involved in mycobacterial virulence. Histological and infection assays showed that the M. marinum mimG mutant, an Rv3242c orthologue in a pathogenic M. marinum strain, was strongly attenuated in adult zebrafish and also survived less in macrophages. In contrast, infection with wild type and the complemented ΔmimG:Rv3242c M. marinum strains showed prominent pathological features, such as severe emaciation, skin lesions, hemorrhaging, and more zebrafish death. Similarly, recombinant MsmRv3242c bacteria showed increased invasion in non-phagocytic epithelial cells and longer intracellular survival in macrophages as compared with wild type and vector control M. smegmatis strains. Further mechanistic studies revealed that the Rv3242c- and mimG-mediated enhancement of intramacrophagic survival was due to inhibition of autophagy, reactive oxygen species, and reduced activities of superoxide dismutase and catalase enzymes. Infection with MsmRv3242c also activated the MAPK pathway, NF-κB, and inflammatory cytokines. In summary, we show that a novel mycobacterial mannosylated phosphoribosyltransferase acts as a virulence and immunomodulatory factor, suggesting that it may constitute a novel target for antimycobacterial drugs.