Analysis of in vitro activity of high dilutions of Euphorbia tirucalli L. in human melanoma cells

Aveloz (firestick cactus; Euphorbia tirucalli L.) belongs to Euphorbiaceae family, characterized by the production of a toxic latex that has corrosive effects on the skin and mucous membranes. Continual topic use of the latex is recommended by popular medicine to treat warts, and epitheliomas. To validate this indication, ultra diluted latex and homeopathic medicine Euphorbia tirucalli were tested in vitro on the proliferation of melanoma cells. Ultra diluted latex was prepared in homeopathic dilutions 5cH, 15cH and 30cH by dilution and agitation (trituration for solid and sucussion for liquid phases) using 70º GL (Gay Lussac) ethylic alcohol (70º GL EtOH 70ºGL) as inert medium according to the guidelines in Farmacopéia Homeopática Brasileira (FHB). Homeopathic medicine Euphorbia tirucalli was prepared from mother-tincture according to the centesimal Hahnemannian method. Solutions 0.5% and 5% of 70ºGL EtOH were succussed and used as control. Human melanoma cells were cultured, treated and monitored by method MTT for 24 to 72 hours. It was observed that 0.5% 70ºGL EtOH solution had little or no effect on the proliferation of melanoma cells (5.1% maximal inhibition in dilution 30cH). Positive correlation was observed in most groups between inhibition of proliferation and diluted preparations, maximal increase (9%) was seen in with 5% latex. Moreover, mother-tincture proved to be more active than latex; treatment with 0.5% solution of latex 30cH exhibited 19.7% inhibition, whereas treatment with 0.5% solution of Euphorbia tirucalli 30cH exhibited 32.1% inhibition of cell proliferation (p

Study on the Anti-cancer Potential of Aegle marmelos Fruit Extract Pro and Anti Apoptotic Molecule in Human Melanoma Cell Line-A375

Aegle marmelos is also known as bael which is commonly found in south East Asia and Indian-sub continent. The origin of bael is India. Bael is also known as the golden apple, Bengal-quince in India. In the ancient medical system, Aegle marmelos play an important role and its extract is also useful in inflammation, diabetes, cancer and asthma. The leaves are used for anti-inflammatory, nervous disorder, control blood sugar and fruit is used to treat antiviral, anti-diabetics, and brain and heart tonic. In addition, studies have proved that bael is used for the treatment and prevention of cancer. The main aim of this study is to assess the anti-cancer potential of Aegle marmelos fruit extract pro and anti apoptotic molecules in human melanoma cell line-A375. In the present study, Human Melanoma A375 cells will be produced, grown and will be passed in different culture flasks, then RNA isolation was done and by reverse transcriptase process, RNA gets converted into cDNA. This cDNA will be used for the amplification of growth factor beta using gene specific primers by commercially available real time PCR kit. The anticancer potential effect was found in 400µg/ml, as the concentration increases, the cell viability is decreased. The current study explains the potential application of bael in pharmacological and medicinal uses in near future.

Apoptosis Pathways Triggered by a Potent Antiproliferative Hybrid Chalcone on Human Melanoma Cells

The World Health Organization reported that approximately 324,000 new cases of melanoma skin cancer were diagnosed worldwide in 2020. The incidence of melanoma has been increasing over the past decades. Targeting apoptotic pathways is a potential therapeutic strategy in the transition to preclinical models and clinical trials. Some naturally occurring products and synthetic derivatives are apoptosis inducers and may represent a realistic option in the fight against the disease. Thus, chalcones have received considerable attention due to their potential cytotoxicity against cancer cells. We have previously reported a chalcone containing an indole and a pyridine heterocyclic rings and an α-bromoacryloylamido radical which displays potent antiproliferative activity against several tumor cell lines. In this study, we report that this chalcone is a potent apoptotic inducer for human melanoma cell lines SK-MEL-1 and MEL-HO. Cell death was associated with mitochondrial cytochrome c release and poly(ADP-ribose) polymerase cleavage and was prevented by a non-specific caspase inhibitor. Using SK-MEL-1 as a model, we found that the mechanism of cell death involves (i) the generation of reactive oxygen species, (ii) activation of the extrinsic and intrinsic apoptotic and mitogen-activated protein kinase pathways, (iii) upregulation of TRAIL, DR4 and DR5, (iv) downregulation of p21Cip1/WAF1 and, inhibition of the NF-κB pathway.

Different Glucose Metabolic Features According to Cancer and Immune Cells in the Tumor Microenvironment

BackgroundA close metabolic interaction between cancer and immune cells in the tumor microenvironment (TME) plays a pivotal role in cancer immunity. Herein, we have comprehensively investigated the glucose metabolic features of the TME at the single-cell level to discover feasible metabolic targets for the tumor immune status.MethodsWe examined expression levels of glucose transporters (GLUTs) in various cancer types using The Cancer Genome Atlas (TCGA) data and single-cell RNA-seq (scRNA-seq) datasets of human cancer tissues including melanoma, head and neck, and breast cancer. In addition, scRNA-seq data of immune cells in the TME acquired from human melanoma after immune checkpoint inhibitors were analyzed to investigate the dynamics of glucose metabolic profiles of specific immune cells.ResultsPan-cancer bulk RNA-seq showed that the GLUT3-to-GLUT1 ratio was positively associated with immune cell enrichment score. The scRNA-seq datasets of various human cancer tissues showed that GLUT1 was highly expressed in cancer cells, while GLUT3 was highly expressed in immune cells in TME. The scRNA-seq data obtained from human melanoma tissues pre- and post-immunotherapy showed that glucose metabolism features of myeloid cells, particularly including GLUTs expression, markedly differed according to treatment response.ConclusionsDifferently expressed GLUTs in TME suggest that GLUT could be a good candidate a surrogate of tumor immune metabolic profiles and a target for adjunctive treatments for immunotherapy.

A Review of Progesterone Effects on Human Melanoma Cell Growth In-Vitro

Progesterone, a female sex hormone not only has a role in reproduction, but also in protecting females in melanoma. A survey of steroid hormones actions steroid hormones actions survey on melanoma cells and literature survey showed that progesterone inhibited mouse and human melanoma cell growth significantly in-vitro. Progesterone not only inhibited cell growth, but also affected adhesion and migration functions (essential for metastasis) in-vitro. This observation correlated with the clinical studies where they had shown showed an increased survival and delayed metastasis in menstruating females in melanoma. Further, progesterone level in menstruating females (1000–1500 ng/dL) compared to post-menopausal females (20–100 ng/dL) also correlated with previous clinical studies. Progesterone action on melanoma cells, as reported by other researchers also supported the findings from this lab. Hence, progesterone could be the steroid hormone protecting menstruating females in melanoma. Moreover, our recent studies showed that progesterone suppressed pro-inflammatory cytokine IL-8 secretion by the melanoma cells, which decreased melanoma cell growth in-vitro. Hence, progesterone apart from reproductive function may also be involved in protecting menstruating females in melanoma.

AKT1 is Required for a Complete Palbociclib-induced Senescence Phenotype in BRAF-V600E-Driven Human Melanoma

Cellular senescence is a carefully regulated process of proliferative arrest accompanied by numerous functional and morphologic changes. Senescence allows damaged cells to avoid neoplastic proliferation, however induction of the senescence-associated secretory phenotype (SASP) can promote tumor growth. The complexity of the senescence response may limit the efficacy of anti-neoplastic agents, such as CDK4/6 inhibitors (Cdk4/6i), that induce a senescence-like, non-proliferative state in tumor cells. The AKT kinase family plays an important role in cellular growth and division, and is commonly hyperactive in many cancers including melanoma. AKT activity has also been implicated in regulation of senescence. The three AKT isoforms play both redundant and unique roles in tumorigenesis and cancer progression. To interrogate the role of AKT isoforms in the induction of cellular senescence by Cdk4/6i, we generated isoform specific AKT knockout human BRAF-V600E mutated melanoma cell lines. We found that the CDK4/6i Palbociclib induced a form of senescence in these cells that was dependent on AKT1. As a potential mechanism, we evaluated the activity of the cGAS-STING pathway, recently implicated in cellular senescence. While we showed cGAS-STING function to be dependent on AKT1, pharmacologic inhibition of either cGAS or STING had little effect on senescence. However, we found SASP factors to require NF-kB function, in part dependent on a stimulatory phosphorylation of IKKα by AKT1 previously reported in other models. In summary, we provide the first evidence of a novel, isoform specific role for AKT1 in therapy-induced senescence in human melanoma cells acting through NF-kB but independent of cGAS-STING.