EEF1A2 promotes HIF1A mediated breast cancer angiogenesis in normoxia and participates in a positive feedback loop with HIF1A in hypoxia.

BACKGROUND: The eukaryotic elongation factor, EEF1A2, has been identified as an oncogene in various solid tumors. Here, we have identified a novel function of EEF1A2 in angiogenesis. METHODS: Chick chorioallantoic membrane, tubulogenesis, aortic ring, Matrigel plug, and skin wound healing assays established EEF1A2's role in angiogenesis. RESULT: Higher EEF1A2 levels in breast cancer cells enhanced cell growth, movement, blood vessel function, and tubule formation in HUVECs, as confirmed by ex-ovo and in-vivo tests. The overexpression of EEF1A2 could be counteracted by Plitidepsin. Under normoxic conditions, EEF1A2 triggered HIF1A expression via ERK-Myc and mTOR signaling in TNBC and ER/PR positive cells. Hypoxia induced the expression of EEF1A2, leading to a positive feedback loop between EEF1A2 and HIF1A. Luciferase assay and EMSA confirmed HIF1A binding on the EEF1A2 promoter, which induced its transcription. RT-PCR and polysome profiling validated that EEF1A2 affected VEGF transcription and translation positively. This led to increased VEGF release from breast cancer cells, activating ERK and PI3K-AKT signaling in endothelial cells. Breast cancer tissues with elevated EEF1A2 showed higher microvessel density. CONCLUSION: EEF1A2 exhibits angiogenic potential in both normoxic and hypoxic conditions, underscoring its dual role in promoting EMT and angiogenesis, rendering it a promising target for cancer therapy.

Regulatory significance of CULLIN2 in neuronal differentiation and regeneration.

BACKGROUND: Scaffold proteins coordinate multiple signalling pathways by integrating various proteins but the role of these proteins in neuronal pathways remains to be elucidated. The present study focused to evaluate the expression of the scaffold protein CULLIN2 in neuronal cells. METHODS: The neuronal precursor cell line N2A was differentiated to neurons in-vitro with retinoic acid and biochemical assays were used to understand the gene expression profiling of CULLIN2. Moreover, neddylation inhibitor MLN4924 was used to inhibit the activity of CULLIN2 and the downstream substrates were validated. Finally, the role of CULLIN2 in nerve regeneration was evaluated in an in vivo zebrafish model. RESULTS: Experimental data showed that the neuronal cells N2A have lower expression of CULLIN2 compared to skin cell lines (HaCaT and A431) and inactivation with the neddylation inhibitor resulted in cell death. Furthermore differentiating the neural precursor cell line into neurons with retinoic acid enhanced the expression of CULLIN2. Examining downstream signalling molecules with the neddylation inhibitor illuminates that MLN4924 treatment influences the cytokine signalling cascade (JAK-STAT) in neuronal cells. Moreover, for the first time, we show that the ubiquitin ligase protein CULLIN2 is perturbed in neural regeneration. Expression profile of CULLIN2 was significantly decreased in response to a nerve injury in Zebra fish and as the nerve regenerates there is corresponding reduction in the mRNA levels. CONCLUSION: During differentiation CULLIN2 is upregulated whereas during regeneration there is significant downregulation. Thus, our findings reveal a crucial role of the scaffold protein CULLIN2 in nerve differentiation and regeneration which can be vital for the treatment of nerve injury.

Applications of molybdenum oxide nanoparticles impregnated collagen scaffolds in wound therapeutics.

INTRODUCTION: The highly complex pathophysiology of the wound micro-environment demands the development of a multi-faceted system which would enhance the wound healing cascade. Incorporation of nanotechnology in wound therapeutics has opened up new avenues to tourment the diseased condition. Amongst the various types of nanoparticles molybdenum oxide nanoparticles posses various inherent properties that makes it a versatile material to be used in healing. Incorporation of Molybdenum nanoparticles into collagen scaffolds would provide a synergistic and sequential healing process ensuring the formation of a fully functional tissue. MATERIALS AND METHODS: The physico-chemical characterization of the synthesized materials were done using SEM and FT-IR techniques. The bicompatibility and cell proliferation were tested using HaCaT cell lines. Pro-angiogenic ability of the scaffold was tested using CAM assay and Chick aortic arch assay. Finally the in-vivo wound healing ability of the material was tested by creating wound of about 6 cm2 on the dorsal side of Wistar rats and observed for about 21 days. RESULTS: The characterization of the scaffold revealed the presence MoO3 nanoparticles and their structural integrity within the scaffold. The synthesized MoO3-collagen nanocomposite was found to be biocompatible and hemocompatible. The in-vitro studies demonstrated that the MoO3-collagen scaffold significantly increased the cell adhesion and migration to nearly 2 fold. The MoO3 embedded collagen sheets synergistically favoured neovascularization and re-epithelization,which would potentially enhance therapeutic efficiency of the scaffold. The nanocomposite also encouraged results in in-vivo analysis, the Wistar rats treated with MoO3-collagen scaffolds showed complete healing in about 15 days. CONCLUSION: The fabricated MoO3-collagen scaffold was found to play an important role in all major events of wound healing such as adhesion, migration, proliferation and angiogenesis. The in-vivo healing assay also proved that the healing rate of animals treated with the samples was comparatively faster. Further research using various trace elements would open up promising avenues in healing therapeutics.

Bi-faceted delivery of phytochemicals through chitosan nanoparticles impregnated nanofibers for cancer therapeutics.

Drug delivery through nanotechnological approaches has predominantly gained significance owing to the enhanced bioavailability, stability and targeted sequel. Multiple drug delivery is also on its stride to achieve holistic therapeutic regime. In our quest for such a treatment for cancer we selected two phytochemicals namely resveratrol (RS) and ferulic acid (FA) that have gained wide attention in the field of medicine due to their array of properties. Albeit their multifaceted application their therapeutic potential is mired due to its physicochemical instability and low bioavailability. Therefore, in the present study combinatorial effect of these compounds in cancer therapeutics have been scrutinized by fabricating chitosan nanoparticle loaded polycaprolactone nanofibers for delivering RS and FA. The materials were physico-chemical characterized. The nanoparticle incorporation within the nanofibers was corroborated through FITC tagging. The anti-cancer effect of drug loaded nanofibers were studied using A431 cells which displayed 30% and 50% reduction in the cell viability when treated with nanoparticles and nanofibrous scaffold. In congruence, the anti-angiogenic potential of the scaffold was elucidated using Chick chorioallantoic membrane assay and aortic ring assay. Thus, the nanofibrous delivery system opens up new venue in the field of cancer therapeutics with reduced side effects and efficient cancer targeting.