Dermaseptin B2’s Anti-Proliferative Activity and down Regulation of Anti-Proliferative, Angiogenic and Metastatic Genes in Rhabdomyosarcoma RD Cells <i>in Vitro</i>

Background: Rhabdomyosarcoma (RMS) is the most prevalent soft tissue sarcoma in children, representing approximately 50% of pediatric sarcomas and can develop in any part of the body though more frequently at the extremities. Aim: Evaluating the in vitro anti-proliferative activity of Dermaseptin B2 on Rhabdomyosarcoma RD (CCL-136TM) cells and its effect on the expression of MYC, FGFR1, NOTCH1, and CXCR7 genes involve in processes including proliferation, angiogenesis and metastasis. Methods: RD cells were grown in Dulbecco’s Modified Eagle’s Medium supplemented with 10% Fetal Bovine Serum. Exponentially growing cells were treated with Dermaseptin B2 and Antiproliferative activity was assayed using the resazurin and migration assays at three time-points. In order to determine the gene expression profiles of MYC, NOTCH1, FGFR1 and CXCR7, total RNA was extracted from the cells and q-RT-PCR was performed with β-Actin as reference gene. Results: Dermaseptin B2 inhibited the proliferation of RD cells in a time and concentration dependent manner as with IC50 values of 7.679 μM, 7.235 μM, 5.993 μM. The 2-dimentional wound healing assay showed inhibition of migration and motility of the RD cells at time-points of 6, 24, 48 and 72-hours with the greatest inhibition observed at 72-hours. Dermaseptin B2 downregulated the target MYC (fc;1.5013, 1.5185, 2.4144), CXCR7 (fc;2.8818, 4.4430, 3.9924), FGFR1 (fc;2.3515, 2.0809, 2.2543), NOTCH1 (fc;2.4667, 4.6274, 4.3352) genes for the three-time points respectively. NOTCH1 and CXCR7 showed higher fold changes with respect to β-Actin than MYC and FGFR1. Conclusion: The results of this study indicate that Dermaseptin B2 is a target molecule for signaling pathways including PI3K/AKT, RTK and NOTCH pathways that could affect the transcription of these genes and overall inhibition of cancer progression. Further studies are needed to give a better understanding of the detailed mechanisms of action as well as the effects of the Dermaseptin B2 peptide in vivo.

Annona muricata:Is the natural therapy to most disease conditions including cancer growing in our backyard?A systematic review of its research history and future prospects

Annona muricata(A.muricata)is a tropical plant species belonging to family Annonaceae and known for its many medicinal uses.This review focuses on the research history of its traditional uses,phytochemicals,pharmacological activities,toxicological aspects of the extracts and isolated compounds,as well as the in vitro propagation studies with the objective of stimulating further studies on this plant for human consumption and treatment.A.muricata extracts have been identified in tropical regions to traditionally treat diverse conditions ranging from fever to diabetes and cancer.More than 200 chemical compounds have been identified and isolated from this plant,the most important being alkaloids,phenols and acetogenins.Using in vitro studies,its extracts and phytochemicals have been characterized as antioxidant,anti-microbial,anti-inflammatory,insecticidal,larvicidal,and cytotoxic to cancer cells.In vivo studies have revealed anxiolytic,antistress,anti-inflammatory,immunomodulatory,antimalarial,antidepressant,gastro protective,wound healing,hepato-protective,hypoglycemic,anticancer and anti-tumoral activities.In silico studies have also been reported.In addition,clinical studies support the hypoglycemic as well as some anticancer activities.Mechanisms of action of some pharmacological activities have been elucidated.However,some phytochemical compounds isolated from A.muricata have shown a neurotoxic effect in vitro and in vivo,and therefore,these crude extracts and isolated compounds need to be further investigated to define the magnitude of the effects,optimal dosage,and mechanisms of action,long-term safety,and potential side effects.Additionally,more clinical studies are necessary to support the therapeutic potential of this plant.Some studies were also found to have successfully regenerated the plant in vitro,but with limited success.The reported toxicity notwithstanding,A.muricata extracts seem to be some of the safest and promising therapeutic agents of the 21st century and beyond that need to be studied further for better medicinal formulations and diseases management.