a375 melanoma
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Author(s):  
Antonina Dadadzhanova ◽  
Ekaterina Kolesova ◽  
Vladimir Maslov ◽  
Eliz Amar-Lewis ◽  
Riki Goldbart ◽  
...  

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1848
Author(s):  
Ewa Mazurkiewicz ◽  
Aleksandra Makowiecka ◽  
Ewa Mrówczyńska ◽  
Iryna Kopernyk ◽  
Dorota Nowak ◽  
...  

Skin melanocytes reside on the basement membrane (BM), which is mainly composed of laminin, collagen type IV, and proteoglycans. For melanoma cells, in order to invade into the skin, melanocytes must cross the BM. It has been reported that changes in the composition of the BM accompany melanocytes tumorigenesis. Previously, we reported high gelsolin (GSN)—an actin-binding protein—levels in melanoma cell lines and GSN’s importance for migration of A375 cells. Here we investigate whether melanoma cells migrate differently depending on the type of fibrous extracellular matrix protein. We obtained A375 melanoma cells deprived of GSN synthesis and tested their migratory properties on laminin, collagens type I and IV, fibronectin, and Matrigel, which resembles the skin’s BM. We applied confocal and structured illuminated microscopy (SIM), gelatin degradation, and diverse motility assays to assess GSN’s influence on parameters associated with cells’ ability to protrude. We show that GSN is important for melanoma cell migration, predominantly on laminin, which is one of the main components of the skin’s BM.


2021 ◽  
Vol 14 (6) ◽  
Author(s):  
Maryam Iranzadasl ◽  
Parvin Pasalar ◽  
Mohammad Kamalinejad ◽  
Mohammad Javad Mousavi

Background: Melanoma is the leading cause of 80% of skin cancer worldwide due to its high proliferation rate, metastatic nature, and limited effective therapies. Given the rapid increase in its incidence compared to other skin cancers, new therapeutic agents are needed to control the disease. Scientists are interested in medicinal plants due to their anticancer properties. The rhizomes of the Iris germanica L., known as “Irsa”, is one of the herbs used in traditional Persian medicine for the treatment of various skin cancers. Objectives: This study aimed at investigating the cytotoxic effects of Iris germanica on A375 melanoma and AGO-1522 normal human fibroblast cell lines for the first time. Methods: The ethanolic extract was prepared by the maceration method. Cell viability and cytotoxic activities were assessed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometric assay, using annexin V/propidium iodide (PI) staining. Results: IC50 values were estimated for the A375 melanoma and the AGO-1522 normal cell lines. We revealed that the IC50 for the A375 melanoma was 0.0438 mg/mL and for the AGO-1522 normal cell line was 0.8494 mg/mL after 48 hours of treatment. Furthermore, flow cytometry analysis illustrated that 0.125 mg/mL of the Iris germanica extract could lead to 55.24% apoptosis of the A375 melanoma cell line. The same concentration of the Iris germanica extracts only lead to 8.76% apoptosis in the AGO-1522 cell line. Conclusions: Iris germanica extract has considerable cytotoxic effects on the human melanoma cell line. Further studies are required to demonstrate the therapeutic effects of Iris germanica on melanoma cancer.


2021 ◽  
Author(s):  
Anindita Mitra ◽  
Rita Ghosh

Abstract Background: Cisplatin has been extensively used in therapeutics for its broad-spectrum anticancer activity and frequently used for the treatment of solid tumors. However, it presents several side-effects and several cancers develop resistance. Combination therapy of cisplatin with poly (ADP-ribose) polymerase 1 (PARP1) inhibitors has been effective in increasing its efficacy at lower doses. Methods and Results: In this work, we have shown that the nitro-flavone derivative, 2-(4-Nitrophenyl)-4H-chromen-4-one (4NCO), can improve the sensitivity of cancer cells to cisplatin through inhibition of PARP1. The effect of 4NCO on cisplatin toxicity was studied through combination therapy in both exponential and density inhibited A375 melanoma cells. Combination index (CI) was determined from isobologram analysis. The mechanism of cell killing was assessed by lactate dehydrogenase (LDH) assay. Temporal nicotinamide adenine dinucleotide (NAD+) assay was done to show the inhibition of PARP1. We also performed in silico molecular modeling studies to know the binding mode of 4NCO to a modeled PARP1-DNA complex containing cisplatin-crosslinked adduct. The results from both in silico and in cellulo studies confirmed that PARP1 inhibition by 4NCO was most effective in sensitizing A375 melanoma cells to cisplatin. Isobologram analysis revealed that 4NCO reduced cell viability both in exponential and density inhibited A375 cells synergistically. The combination led to cell death through apoptosis. Conclusion: The synthetic nitro-flavone derivative 4NCO effectively inhibited the important nuclear DNA repair enzyme PARP1 and therefore, could complement the DNA-damaging anticancer drug cisplatin in A375 cells and thus, could act as a potential adjuvant to cisplatin in melanoma therapy.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Michele Lai ◽  
Rachele Amato ◽  
Veronica La Rocca ◽  
Mesut Bilgin ◽  
Giulia Freer ◽  
...  

AbstractAcid ceramidase (AC) is a lysosomal hydrolase encoded by the ASAH1 gene, which cleaves ceramides into sphingosine and fatty acid. AC is expressed at high levels in most human melanoma cell lines and may confer resistance against chemotherapeutic agents. One such agent, doxorubicin, was shown to increase ceramide levels in melanoma cells. Ceramides contribute to the regulation of autophagy and apoptosis. Here we investigated the impact of AC ablation via CRISPR-Cas9 gene editing on the response of A375 melanoma cells to doxorubicin. We found that doxorubicin activates the autophagic response in wild-type A375 cells, which effectively resist apoptotic cell death. In striking contrast, doxorubicin fails to stimulate autophagy in A375 AC-null cells, which rapidly undergo apoptosis when exposed to the drug. The present work highlights changes that affect melanoma cells during incubation with doxorubicin, in A375 melanoma cells lacking AC. We found that the remarkable reduction in recovery rate after doxorubicin treatment is strictly associated with the impairment of autophagy, that forces the AC-inhibited cells into apoptotic path.


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