Synthesis and photodynamic properties of pyrazole-indole hybrids in the human skin melanoma cell line G361

Summary Randia dumetorum (family Rubiaceae) is highly reputed ayurvedic medicinal tree commonly known as the Mainphal. A large deciduous thorny shrub grows up to 5 m of height. It occurs almost throughout India up to 1200 m of altitude. It is found in Himalaya from Jammu East ward ascending to 400 m and from Kashmir to East ward up to 1200 m. 11-methylixoside (compound 1), an iridoid glucoside, was isolated from the bark of this plant. The structure was characterized by using spectroscopic methods including 1D-1HNMR,13C-NMR and 2D-NMR (HSQC,HMBC, DQF-COSY) experiments and confirmed by comparison of their NMR data with those from the literature. This compound has been reported for the first time in Randia dumetorum bark. The 11-methylixoside was subjected to cytotoxic activity against MDA-MB-231 (breast cancer cell line) and SK-MEL-2 (human skin melanoma cell line), BE(2)C (neuroblastoma cell line derived from human bone marrow) and U87MG (human neuronale glioblastoma (astrozytom) cell line showed appreciable cytotoxic effect with IC50 value 63.10 µg/ml concentration for SK-MEL-2 (human skin melanoma cell line).

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CHARACTERISTICS OF MULTIDRUG RESISTANCE IN HUMAN SKIN MELANOMA CELL LINES

Russian Journal of Biotherapy ◽

10.17650/1726-9784-2015-14-4-39-44 ◽

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.

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Quantitative Autofluorescence Imaging of A375 Human Melanoma Cell Samples: A Pilot Study

Journal of lasers in medical sciences ◽

10.34172/jlms.2021.04 ◽

2021 ◽

Vol 12 ◽

pp. e4-e4

Author(s):

Afshan Shirkavand ◽

Ezeddin Mohajerani ◽

Shirin Farivar ◽

Leila Ataie-Fashtami ◽

Mohammad Hossein Ghazimoradi

Keyword(s):

Image Processing ◽

Pilot Study ◽

Optical Imaging ◽

Cell Line ◽

Human Skin ◽

Melanoma Cell ◽

Melanoma Cell Line ◽

Human Melanoma ◽

Human Melanoma Cell ◽

Endogenous Fluorophores

Introduction: Skin cancer is one of the most common types of malignancy worldwide. Human skin naturally contains several endogenous fluorophores, as potential sources can emit inherent fluorescence, called intrinsic autofluorescence (AF). The melanin endogenous fluorophore in the basal cell layer of the epidermis seems to have a strong autofluorescence signal among other ones in the skin. This pilot study aimed to investigate the feasibility of the detection of autofluorescence signals in the A375 human melanoma cell line in the cell culture stage using the FluoVision optical imaging system. Methods: The human skin melanoma cell line (A375) donated as a gift from Switzerland (University Hospital Basel) was cultured. For the imaging of the A375 human melanoma cell sample in this pilot study, the FluoVision optical imaging device (Tajhiz Afarinan Noori Parseh Co) was applied. The proposed clustering image processing code was developed based on the K-mean segmentation method, using MATLAB software (version 16). Results: The quantification of color pixels in the color bar along with the intensity score of the autofluorescence signal ranged between 0 and 70 was written in the image processing code execution and a threshold higher than 40%, proportional to the ratio of autofluorescent cells. The percentage of the signal of A375 autofluorescent melanoma cells in the 3 studied cell samples was calculated as 3.11%±0.6. Conclusion: This imaging method has the advantage of no need for fluorophore labels over the existing fluorescence imaging methods, and it can be regarded as one of the important choices of label-free imaging for this A375 melanoma cell line containing the intrinsic endogenous fluorophore in cell studies.

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MODELING OF A SUBCUTANEOUS XENOGRAFT OF HUMAN SKIN MELANOMA MEL CHER WITH V600E BRAF MUTATION IN IMMUNODEFICIENT MICE FOR PRECLINICAL STUDY THE TARGETING ANTICANCER DRUGS

Russian Journal of Biotherapy ◽

10.17650/1726-9784-2016-15-4-65-71 ◽

2016 ◽

Vol 15 (4) ◽

pp. 65-71 ◽

Cited By ~ 1

Author(s):

N. V. Andronova ◽

L. F. Morozova ◽

I. N. Mikhailova ◽

A. A. Lushnikova ◽

D. A. Ponkratova ◽

...

Keyword(s):

Cancer Research ◽

Complete Remission ◽

Cell Line ◽

Melanoma Cell ◽

Braf Mutation ◽

Melanoma Cell Line ◽

Target Therapy ◽

Molecular Genetic ◽

Skin Melanoma

Introduction. Development of new models of a human disseminated skin melanoma of the with molecular-genetic targets for specific therapy increases productivity of the preclinical researches new the anti-melanoma drugs or their combinations in vitro and in vivo. Such opportunity is realized by adaptation in vivo of the original human pigmented skin melanoma cell line mel Cher and receiving subcutaneous (s. c.) xenograft under monitoring of transplant, morphological, molecular-genetic (V600E BRAF mutation) and chemotherapeutic (sensitivity for the inhibitor of BRAF kinases to a vemurafenib) characteristics. Objective: receiving from the cell line mel Cher s. c. xenogratft of the human pigmented skin melanoma with V600E BRAF mutation and sensitive to specific target therapy. Materials and methods. Human pigmented skin melanoma cell line mel Cher from the Collection of Russian Cancer Research Center and immunodeficient Balb/c nude female mice cultivated in Russian Cancer Research Center was used. Required characteristics are defined by multiple s. c. transplanting in vivo by methods of transplant biology, a light microscopy, molecular-genetics and the experimental chemotherapy. Sensitivity to a BRAF kinase inhibitor to a vemurafenib was estimated under monitoring of the tumor growth rate (Vt/V0) on indexes, adequate for patients: existence of the complete remission and possibility of recurrence. Results. When s. c. transplantation of 107 cell of mel Cher line cytological identical intertwined s. c. xenografts with a stable growth kinetics on 4-9 passages (a latent phase 8 days, exponential - to 14 days, stationary - to 24 days) and existence of a mutation of V600E BRAF have been recieved. Vemurafenib in a single dose of 75 mg/kg caused the complete remission during a 15-day course and within 7 days after its cancellation - with the subsequent recurrence. Conclusion: receiving from the cell line mel Cher s. c. xenogratft of a human pigmented melanoma of skin with a mutation of V600E BRAF and sensitive to specific target therapy is suitable for preclinical studying of the new anti-melanoma drugs specific for this target.

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Monitoring the Response of Skin Melanoma Cell Line (A375) to Treatment with Vemurafenib: A Pilot In Vitro Optical Spectroscopic Study

Photobiomodulation Photomedicine and Laser Surgery ◽

10.1089/photob.2020.4887 ◽

2021 ◽

Vol 39 (3) ◽

pp. 164-177

Author(s):

Afshan Shirkavand ◽

Ezeddin Mohajerani ◽

Shirin Farivar ◽

Leila Ataie-Fashtami ◽

Mohammad Hossein Ghazimoradi

Keyword(s):

Spectroscopic Study ◽

Cell Line ◽

Melanoma Cell ◽

Melanoma Cell Line ◽

Skin Melanoma

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Insight into Mechanistic Action of Thymoquinone Induced Melanogenesis in Cultured Melanocytes

Protein and Peptide Letters ◽

10.2174/0929866526666190506114604 ◽

2019 ◽

Vol 26 (12) ◽

pp. 910-918

Author(s):

Kamal U. Zaidi ◽

Firoz N. Khan ◽

Sharique A. Ali ◽

Kausar P. Khan

Keyword(s):

Western Blot ◽

Signaling Pathways ◽

Cell Line ◽

Melanoma Cell ◽

Melanoma Cell Line ◽

Protein Kinase Inhibitors ◽

Tyrosinase Activity ◽

Dependent Manner ◽

B16f10 Melanoma ◽

B16f10 Melanoma Cell

Background: Melanin plays a crucial role in camouflage, social communication and protection against harmful ultraviolet radiations. Melanin is synthesized by melanocytes through melanogenesis and several intrinsic and extrinsic factors are involved during the process. Any change occuring in the normal melanogenesis process can cause severe pigmentation problems of hypopigmentation or hyperpigmentation. Objective: The present study is based on the evaluation of the effect of thymoquinone on melanogenesis and their possible mechanism of action using the B16F10 melanoma cell line for the production via blocking signaling pathways. Methods: Phase contrast microscopy, cell viability, tyrosinase activity, melanin content and western blot analysis were used in the present study. Results: In the present investigation, cultured melanocytes exhibit that the stimulation of melanin synthesis when treated with thymoquinone. Tyrosinase activity and melanin production in B16F10 melanoma cell line was increased in doze-dependent manner. In western blot, we investigated the involvement of the cAMP/PKA pathway in thymoquinone induced melanogenesis. It was observed protein kinase inhibitors PKA, PKC, PKB and MEK1 decreased the stimulatory effects of thymoquinone from 11.45- fold value to 8.312, 6.631, 4.51, and 7.211-fold value, respectively. However, the results also prove that thymoquinone may partially induce tyrosinase expression via PKA, PKB, PKC and MEK1 signaling pathways. Conclusion: The present finding proposed that thymoquinone is a protective challenger for melanogenesis and it might be useful for the treatment of hypopigmentary disorders.

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In Vitro Anticancer Potential of Jasione montana and Its Main Components Against Human Amelanotic Melanoma Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22073345 ◽

2021 ◽

Vol 22 (7) ◽

pp. 3345

Author(s):

Aleksandra Maria Juszczak ◽

Robert Czarnomysy ◽

Jakub Władysław Strawa ◽

Marijana Zovko Končić ◽

Krzysztof Bielawski ◽

...

Keyword(s):

Cell Line ◽

Melanoma Cell ◽

Melanoma Cell Line ◽

Binding Assay ◽

Annexin V ◽

Melanoma Cells ◽

Amelanotic Melanoma ◽

Cytotoxic Activities ◽

Ionization Mass ◽

Caspase 9

Jasione montana L. (Campanulaceae) is used in traditional Belarusian herbal medicine for sleep disorders in children, but the chemical composition and biological activity have not been investigated. In this study, the activities of J. montana extracts, their fractions and main compounds were evaluated in amelanotic melanoma C32 (CRL-1585) cells and normal fibroblasts (PCS-201-012). The extracts and fractions were analyzed using liquid chromatography–photodiode array detection–electrospray ionization–mass spectrometry (LC–PDA–ESI–MS/TOF) to characterize 25 compounds. Further, three major and known constituents, luteolin (22) and its derivatives such as 7-O-glucoside (12) and 7-O-sambubioside (9) were isolated and identified. The cytotoxic activities against fibroblasts and the amelanotic melanoma cell line were determined using the fixable viability stain (FVS) assay. The influence of diethyl ether (Et2O) fraction (JM4) and 22 on apoptosis induction was investigated using an annexin V binding assay. The obtained results showed significant cytotoxicity of JM4 and 22 with IC50 values of 119.7 ± 3.2 and 95.1 ± 7.2 μg/mL, respectively. The proapoptotic potential after 22 treatment in the C32 human amelanotic melanoma cell line was comparable to that of vinblastine sulfate (VLB), detecting 29.2 ± 3.0% apoptotic cells. Moreover, 22 displayed less necrotic potential against melanoma cells than VLB. In addition, the influences of JM4 and 22 on the dysfunction of the mitochondrial membrane potential (MMP), cell cycle and activity of caspases 3, 8, 9, and 10 were established. The effects of JM4 on MMP change (74.5 ± 3.0% of the cells showed a reduced MMP) corresponded to the results obtained from the annexin V binding assay and activation of caspase-9. JM4 and 22 displayed a significant impact on caspase-9 (40.9 ± 2.4% of the cells contained active caspase-9 after JM4 treatment and 16.6 ± 0.8% after incubation with 22) and the intrinsic (mitochondrial) apoptotic pathway. Moreover, studies have shown that JM4 and 22 affect the activation of external apoptosis pathways by inducing the caspase-8 and caspase-10 cascades. Thus, activation of caspase-3 and DNA damage via external and internal apoptotic pathways were observed after treatment with JM4 and 22. The obtained results suggest that J. montana extracts could be developed as new topical preparations with potential anticancer properties due to their promising cytotoxic and proapoptotic potential.

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