scholarly journals Preliminary Results of Betulinic Acid-Loaded Magnetoliposomes - a Potential Approach to Increase Therapeutic Efficacy in Melanoma

2019 ◽  
Vol 70 (9) ◽  
pp. 3372-3377 ◽  
Author(s):  
Claudia Geanina Farcas ◽  
Elena Alina Moaca ◽  
Razvan Dragoi ◽  
Delia Berceanu Vaduva ◽  
Iasmina Marcovici ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Betulinic Acid ◽  
Biomedical Applications ◽  
Preliminary Evidence ◽  
Structure Preserving ◽  
Healthy Cell ◽  
Magnetic Features ◽  
Melanoma Cell Lines

Magnetoliposomes were placed into the biomedical spotlight due to their unique structure, preserving also magnetic features valuable for biomedical applications. The present study provides a preliminary evidence of the cytotoxicity induced by betulinic acid-loaded magnetoliposomes (BA-Fe3O4@Lip) on two melanoma cell lines - A375 and B164A5 cells and one healthy cell line - mice epidermis JB6 Cl 41-5a cells, augmented by references to potential biomedical applications. BA-Fe3O4@Lip showed significant cytotoxicity on the melanoma cell lines, compared with the blank liposomal structures. In addition, the healthy cell line displayed good viability rate after exposure to BA-Fe3O4@Lip.

scholarly journals CHARACTERISTICS OF MULTIDRUG RESISTANCE IN HUMAN SKIN MELANOMA CELL LINES

2015 ◽  
Vol 14 (4) ◽  
pp. 39-44
Author(s):  
D. A. Afanasieva ◽  
M. A. Baryshnikova ◽  
T. N. Zabotina ◽  
A. A. Borunova ◽  
O. S. Burova ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Cell Line ◽  
Human Skin ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Mdr1 Gene ◽  
Multi Drug Resistance ◽  
Cytotoxic Action ◽  
Skin Melanoma ◽  
Melanoma Cell Lines

MDR is the main obstacle to chemotherapy efficiency. MDR can grow in cancer cells even if only the one cytostatic agent will act. The aim of the nowadays work is to characterize MDR in metastatic human skin melanoma cell lines prepared in “N.N. Blokhin Russian Cancer Research Center”. pgpl70 expression was detected by immunofluorescence methods. mRNA of MDR gene was identified by Reverse Transcriptase- PCR( RT-PCR) method. Rhodamine 123 (Rhl23) emission has been evaluated by flow cyto- fluorimetiy, cytotoxic activity was estimated by MTT-tests. The cells sensitivity to Aianoza cytostatic effects has showed that mel Kor cells were sensitive to Aranoza acting, but mel Ibr and mel Mtp X were not. Mel Ibr cells had expressed pgpl70 from 35 to 50 per cent, it was detected by immunofluorescence reaction. Mel Kor and mel Mtp-X cells were not expressed P-glycoprotein. mRNA of genes responsible for multi-drug resistance - MDR1, BCRP, MRP1 and LRP (MVP) - were detected by PCR. mRNA of BCRP and MRP1 genes has low expression, barely visible stripes after 33 cycles in all cell lines samples. LRP (MVP) genes expression of mRNA, unfortunately, never managed to see. YB1 gene mRNA expression is well, it is typically for cancer cells. mRNA of gene was found in mel MtpX and mel Ibr subclones cell lines. Mel Kor cells didn't contain mRNA of MDR1 gene. The study of the Rhl23 emission from cells showed that mel Kor control cells had accumulated Rhl23 and didn't throw it out. Mel Ibr cell line accumulated Rhl23 and threw out the half part of it. Mel MtpX cell tine had accumulated the less part of Rhl23 and almost all were thrown out. Thus, the study shows that mel Kor cell tine that are sensitive to Aranoza doesn't express pgpl70, not contain mRNA of multi-chug resistance genes and does not throw Rhl23. Mel Ibr cells resistant to the Aranoza cytotoxic action express pgpl70 ,contain mRNA of MDR1 gene and throw out Rhl23. However, mel MtpX cell line resistant to Aranoza does not express pgpl70, but contains mRNA of MDR1 gene and actively throws out Rhl23.

scholarly journals Standard melanoma-associated markers do not identify the MM127 metastatic melanoma cell line

2015 ◽  
Author(s):  
Parvathi Haridas ◽  
Jacqui A. McGovern ◽  
Ahishek S. Kashyap ◽  
D.L. Sean McElwain ◽  
Matthew Simpson
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Protein Level ◽  
Melanoma Cell Line ◽  
Reliable Identification ◽  
Metastatic Melanoma Cell ◽  
Metastatic Cell Line ◽  
Melanoma Cell Lines ◽  
Hmb 45

AbstractReliable identification of different melanoma cell lines is important for many aspects of melanoma research. Common markers used to identify melanoma cell lines include: S100; HMB-45; and Melan-A. We explore the expression of these three markers in four different melanoma cell lines: WM35; WM793; SK-MEL-28; and MM127. The expression of these markers is examined at both the mRNA and protein level. Our results show that the metastatic cell line, MM127, cannot be detected using any of the commonly used melanoma-associated markers. This implies that it would be very difficult to identify this particular cell line in a heterogeneous sample, and as a result this cell line should be used with care.

scholarly journals Cytotoxic Effect of Vanicosides A and B from Reynoutria sachalinensis against Melanotic and Amelanotic Melanoma Cell Lines and in silico Evaluation for Inhibition of BRAFV600E and MEK1

2020 ◽  
Vol 21 (13) ◽  
pp. 4611 ◽  
Author(s):  
Izabela Nawrot-Hadzik ◽  
Anna Choromańska ◽  
Renata Abel ◽  
Robert Preissner ◽  
Jolanta Saczko ◽  
...  
Keyword(s):  
Cell Death ◽  
Molecular Docking ◽  
Cell Line ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Primary Fibroblast ◽  
Fibroblast Line ◽  
Reynoutria Sachalinensis ◽  
A375 Cell Line ◽  
Melanoma Cell Lines

Vanicosides A and B are the esters of hydroxycinnamic acids with sucrose, occurring in a few plant species from the Polygonaceae family. So far, vanicosides A and B have not been evaluated for anticancer activity against human malignant melanoma. In this study, we tested these two natural products, isolated from Reynoutria sachalinensis rhizomes, against two human melanoma cell lines (amelanotic C32 cell line and melanotic A375 cell line, both bearing endogenous BRAFV600E mutation) and two normal human cell lines—keratinocytes (HaCaT) and the primary fibroblast line. Additionally, a molecular docking of vanicoside A and vanicoside B with selected targets involved in melanoma progression was performed. Cell viability was studied using an MTT assay. A RealTime-Glo™ Annexin V Apoptosis and Necrosis assay was used for monitoring programmed cell death (PCD). Vanicoside A demonstrated strong cytotoxicity against the amelanotic C32 cell line (viability of the C32 cell line was decreased to 55% after 72 h incubation with 5.0 µM of vanicoside A), significantly stronger than vanicoside B. This stronger cytotoxic activity can be attributed to an additional acetyl group in vanicoside A. No significant differences in the cytotoxicity of vanicosides were observed against the less sensitive A375 cell line. Moreover, vanicosides caused the death of melanoma cells at concentrations from 2.5 to 50 µM, without harming the primary fibroblast line. The keratinocyte cell line (HaCaT) was more sensitive to vanicosides than fibroblasts, showing a clear decrease in viability after incubation with 25 µM of vanicoside A as well as a significant phosphatidylserine (PS) exposure, but without a measurable cell death-associated fluorescence. Vanicosides induced an apoptotic death pathway in melanoma cell lines, but because of the initial loss of cell membrane integrity, an additional cell death mechanism might be involved like permeability transition pore (PTP)-mediated necrosis that needs to be explored in the future. Molecular docking indicated that both compounds bind to the active site of the BRAFV600E kinase and MEK-1 kinase; further experiments on their specific inhibitory activity of these targets should be considered.

scholarly journals Synthesis and In Vitro Antiproliferative Activity of New -Phenyl-3-(4-(Pyridin-3-Yl)Phenyl)Urea-Scaffold Based Compounds

2017 ◽  
Author(s):  
Mohammad M. Al-Sanea ◽  
So Ha Lee ◽  
Mohammed Safwan Ali Khan ◽  
Pooi-Ling Mok ◽  
Bahaa G. M. Youssif
Keyword(s):  
Breast Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Melanoma Cell ◽  
Antiproliferative Activity ◽  
Cancer Cell Lines ◽  
Cancer Types ◽  
Melanoma Cell Lines

Synthesis of new series of 1-phenyl-3-(4-(pyridin-3-yl) phenyl) urea derivatives and its in vitro antiproliferative activities against NCI-60 human cancer cell lines of nine different cancer types are described. Fourteen compounds 5a-n have been synthesized with three different hydrogen bondable moieties (4-hydroxylmethylpiperidinyl and trimethoxyphenyloxy and 4-hydroxyethylpiperazine) attached to the core structure 1-phenyl-3-(4-(pyridin-3-yl) phenyl) urea. Different substituents with different π and σ values were added on the terminal phenyl group. Compounds with 4-hydroxymethylpiperidine moiety showed higher mean percentage inhibition values over the 60-cell line panel at 10-µM concentration. They showed broad-spectrum antiproliferative activity over many cell lines of different cancer types. For instance, compound 5a elicited some lethal rather than inhibition effects on SK-MEL-5 melanoma cell line, 786-0, A498, RXF 393 renal cancer cell lines, and MDA-MB-468 breast cancer cell line by 146.1, 108.7, 136.2, 134.8, 116.6 % at 10 µM, respectively. Compounds 5a-e exhibited superior antiproliferative activity than Paclitaxel and Gefitinib against the most sensitive cell lines. Two compounds, 5a and 5d showed promising mean growth inhibitions and thus were further tested at five-dose testing mode to determine their IC50 values. The data revealed that 5a and 5d urea compounds are the most active derivatives with significant efficacies and superior potencies than Paclitaxel in 21 different cancer cell lines, belonging particularly to renal cancer and melanoma cell lines. Moreover, 5a and 5d had superior potencies than Gefitinib in 38 and 34 cancer cell lines, respectively; belonging particularly to colon cancer, breast cancer and melanoma cell lines.

Selective activity of dasatinib for the most common KIT mutation in melanoma (L576P)

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 9019-9019
Author(s):  
S. E. Woodman ◽  
J. C. Trent ◽  
K. Stemke-Hale ◽  
A. Lazar ◽  
S. Pricl ◽  
...  
Keyword(s):  
Molecular Modeling ◽  
Cell Line ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Human Melanoma ◽  
Mucosal Melanoma ◽  
Kit Mutation ◽  
Dasatinib Treatment ◽  
Melanoma Cell Lines

9019 Background: Point mutations in the KIT receptor tyrosine kinase gene have recently been identified in melanomas from mucosal, acral lentiginous (AL), and chronically sun-damaged (CSD) sites. An improved understanding of the molecular characteristics of melanoma-prevalent KIT mutations may lead to more effective therapeutic approaches. Methods: Human melanoma cell lines were screened by mass-spectroscopy based genotyping for KIT mutations, and a cell line with an endogenous L576P KIT mutation was identified. The cell line was treated in vitro with a panel of small molecule KIT inhibitors and the effects on cell viability were quantified. Molecular modeling of the interaction of the inhibitors with the KIT L576P mutant protein was determined to estimate binding affinity. PET/CT studies were performed on patients with mucosal melanoma harboring the L576P mutation pre- and post-dasatinib treatment. Results: We have identified the first human melanoma cell line with an endogenous L576P mutation, the most common KIT mutation in melanoma. In vitro testing demonstrated that this cell line is resistant to imatinib, nilotinib and sorafenib (0 - 1 uM), KIT inhibitors shown to be effective in non-melanoma cells with other KIT mutations. However, the mutant cell line was inhibited by dasatinib at concentrations as low as 10 nM, and was significantly more sensitive than melanoma cell lines with wild-type KIT (p = 0.02). No difference in sensitivity to Src inhibitors was observed, supporting that this sensitivity was due to KIT inhibition. Molecular modeling demonstrated that the L576P mutation induces structural changes in KIT that reduce the affinity for imatinib but not for dasatinib. Two metastatic melanoma patients with the L576P KIT mutation were treated with dasatinib, including one patient previously treated with imatinib. Both patients had marked reduction and elimination of tumor FDG- avidity by PET imaging after dasatinib treatment. Conclusions: This data supports that dasatinib has a selective inhibitory effect against the most common KITmutation in melanoma and has implications for the development of AL, CSD, and mucosal melanoma treatment. [Table: see text]

Molecular profiling and comparative genomic hybridization analysis in human melanoma cell lines and identification of BRAF amplification in a PLX4032 acquired resistance cell line.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 8592-8592
Author(s):  
Erika Maria Von Euw ◽  
Judy Dering ◽  
Lee Anderson ◽  
Charles Ginther ◽  
Hsiao-Wang Cheng ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Acquired Resistance ◽  
Braf Inhibitor ◽  
Gene Copy ◽  
Comparative Genomic ◽  
Parental Cell Line ◽  
Molecular Heterogeneity ◽  
Melanoma Cell Lines

8592 Background: Melanoma is the most aggressive form of skin cancer. Its management is evolving rapidly due to an improved understanding of the molecular heterogeneity and the development of effective, personalized targeted therapies. PLX4032 is a BRAF inhibitor that induces tumor response in patients with BRAF mutation whose response is limited due to acquired resistance. We used comprehensive microarray and genomic analysis to molecularly characterize a panel of melanoma cell lines with the goal of identifying new molecular subgroups. To better understand the mechanisms of acquired resistance to PLX4032, we included two cell lines that were conditioned in vitro to acquire resistance. Methods: Using microarray, we studied 52 melanoma cell lines for mRNA expression and performed array CGH using genome microarrays. Data analysis was done using Rosetta Resolver and Agilent Analytics 4.0 software. Results: Microarray analysis suggests melanoma is comprised of at least two distinct molecular groups. One group has a differentiated melanocyte phenotype and the other has an expression signature characteristic of progenitor-like cells. The progenitor-like group expresses SOX9, WNT5A and WNT5B and also has the lowest relative expression of MITF. The most differentiated group had the highest MITF and SOX5 levels, while the progenitor-like cell lines have decreased expression. MITF appears to be a strong candidate marker for this group. Positive correlation exists between MITF expression levels and gene copy number, as almost all of the MITF samples amplified by CHG analysis are also overexpressed. A third group shows strong expression of SOX5, SOX10 and Nestin. One of the most important findings is M14-R cell line, conditioned to acquire PLX4032 resistance, has a focal, high level amplification of BRAF (Log2 ratio = 3, 8-fold amplification) when compared with the parental cell line, which is extremely sensitive to PLX4032. Conclusions: These results afford new insight into the potential pathogenesis and classification and provide a greater understanding of melanoma. BRAF amplification could be a novel mechanism of resistance to PLX4032.

An in vitro comparison of the cytotoxicity of sulphur mustard in melanoma and keratinocyte cell lines

2001 ◽  
Vol 20 (9) ◽  
pp. 483-490 ◽  
Author(s):  
C N Smith ◽  
C D Lindsay ◽  
J L Hambrook
Keyword(s):  
Dna Repair ◽  
Cell Line ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Dna Lesions ◽  
Biochemical Basis ◽  
Sulphur Mustard ◽  
Keratinocyte Cell ◽  
Melanoma Cell Lines

In vivo, the pigment producing melanocytes are the most susceptible cell type to sulphur mustard (HD) in the epidermal region of pig skin. It has been postulated that this is due to the melanogenic pathway producing a cytotoxic, free radical cascade within the melanocyte following HD poisoning, leading to cellular necrosis and subsequent inflammation. To test this hypothesis, the cytotoxicity of HD was tested in three human melanoma cell lines and compared to SVK-14 human keratinocytes, a cell line in which the response to HD has already been characterised. The results of both neutral red (NR) and gentian violet (GV) assays showed that all three melanoma cell lines, particularly the G361 line, were less susceptible to the toxic effects of HD than the SVK-14 keratinocyte cell line. Preliminary data indicate that the expression level of the DNA repair cofactor, proliferating cell nuclear antigen (PCNA), is up to 13-fold greater in the HD-resistant cell line G361 compared to the HD-sensitive SVK-14 cell line. The data point to the importance of DNA lesions in HD-induced cell death and to potential mechanisms associated with increased resistance to HD. A dose–response study was carried out to confirm the differences between these two cell lines. It was found that the G361 line is 5-fold more resistant to HD and 5.5-fold more resistant to the cytotoxic effects of H2O2 than the SVK-14 line, as determined by the MTT assay. The results suggest that differences in the relative efficiency of DNA repair processes may underlie these responses. Whilst the study indicates the limitations of using melanoma cell lines (in vitro) to model melanocyte responses to HD, analysis of the biochemical basis of the observed differences in sensitivity to HD could assist in the identification of novel therapeutic strategies against HD.

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