Our a375 cells are available with a variety of different reporters, including the human sodium iodide symporter hnis, firefly. Conventional therapeutic agents are largely unsatisfactory into the treatment of malignant melanoma. Melanoma cells are routinely identified using antigens encoded by genes that are commonly expressed in melanocytes and melanoma cell lines urosevic et al. Human malignant melanoma has poor prognosis because of resistance to apoptosis and therapy. A total of 1,558 genes were filtered out as differentially expressed among the cell lines based on the following criteria. All related inquiries and orders should be sent via email to. Human melanoma cell lines cls cell lines service gmbh.
The blocking of ctla4 using human monoclonal antibodies, such as ipilimumab, is currently used to relieve ctla4mediated inhibition of antitumor immune response in metastatic melanoma. The lack of braf, nras, kit, pi3k, and akt mutations reveal gnaq and gna11 uveal melanoma cells to be distinct among melanoma types. There are many melanoma cell lines that are commercially available for any number of in vitro studies. Two types of human malignant melanoma cell lines revealed. Our study might be useful to develop more rational sema 3amediated therapeutic strategy for the next generation of cancer management. Description ht144 is a malignant human melanoma cell line that displays aneuploid fibroblastic morphology and grows in adherent tissue culture. Seven out of 10 brafv600e mutant cell lines displayed. Ctla4 cytotoxic t lymphocyte antigen4 is traditionally known as a negative regulator of t cell activation. This new regulatory mechanism may provide an explanation of why a subset of melanoma cell lines are exquisitely sensitive to mek inhibition. Characterization of human melanoma cell lines and melanocytes by. The data confirms the mutually exclusive nature of gnaq and gna11 mutations in vitro.
This study represents the most comprehensive molecular analysis of uveal melanoma cell lines performed to date. Patient derived tumor models and melanoma cell lines to date, a number of tumor model systems have been developed and are commonly used in the preclinical studies of the disease. Immunohistochemical ihc staining performed on 20 hnmmm tissue samples demonstrated expression of ipsc markers oct4, sox2, klf4, and cmyc in all. Three of the melanoma cell lines released an activity that inhibited binding of. Recently, an innovative approach based on inhibitors of the mutated braf gene which represents the most prevalent alteration in melanoma patients appears very promising from the clinical point of view.
Melanomas typically occur in the skin, but may rarely occur in the mouth, intestines, or eye uveal melanoma. The human melanoma cell lines m244, m263, and m381 were infected with a lentiviral vector carrying the pdl1 promoter driving a polycistronic reporter cassette expressing both a dsredexpressdr protein and firefly luciferase linked by a 2a picornavirus sequence figure 2a. Cell culture protocol for patientderived melanoma tumor. In women, they most commonly occur on the legs, while in men they are most common on the back. Mouse cell line developed to fasttrack research in braf melanoma. Metastatic melanoma cancer cell panel atcc tcp1014. Apr 20, 2010 blocking oncogenic signaling induced by the brafv600e mutation is a promising approach for melanoma treatment.
B16f10 cell line is a classical melanoma cell line widely employed in pigmentation studies, because it expresses a wild type mc1r, with an intact transduction machinery, which is activated in response to receptor stimulation. Knocking out plaur, a uparencoding gene, in a375p, a375m6 and hct116, which are two human melanoma and a colon carcinoma, respectively, we have observed an increased number of mitochondria in the two melanoma cell lines, while we evidenced an immature biogenesis of mitochondria in the colon carcinoma culture. Jci insight tumor cell oxidative metabolism as a barrier to. Two types of human malignant melanoma cell lines revealed by. However, the distribution of the expression was highly variable among the cell lines under investigation table 3. Blocking oncogenic signaling induced by the brafv600e mutation is a promising approach for melanoma treatment. Scatter blots of viability assays, organized in cell lines and treatment times, with 1 mugmel1, 2 mugmel2, 3 sbcl2, 4 wm164, and 5 wm793. Cell lines the pig1 human normal melanocyte cell line was kindly provided by the department of ophthalmology, peking university third hospital. Standard melanomaassociated markers do not identify the. Human melanoma cell lines of primary and metastatic origin. Jan 14, 20 using a panel of melanoma cell lines derived from the establishment of excised primary and metastatic tumours, we have investigated the ability of dabrafenib to both exert an antiproliferative activity on cultured melanoma cells and block the erk signalling induced by the mutated braf. Malme3m cells form tumors when injected into immunocompromised mice. Get a printable copy pdf file of the complete article 1.
Melanoma cell lines are an essential tool for melanoma research yu et al. Gene expression profiles of melanoma cells with differing invasivemetastatic potentials. Origin and general characteristics name of cell line cell type organism, ethnicity age gender tissue primary tumor morphology growth properties cls order no. Anastopoulos,1,16 clemenskrepler,2 patriciabrafford,2 katrinsproesser,2 yuchaojiang,3 bradley wubbenhorst,1 ravi amaravadi,4,5 joseph bennett,6 marilda beqiri,2 david elder,5,8 keith t. Three braf mutated melanoma cell lines, a375, igr37 and 501mel each one in its parental p, vemurafenib xp and dabrafenib gpresistant form were treated with 2 concentrations 1 and 10. Combination therapy with vemurafenib plx4032rg7204 and. Collection of three canine melanoma cell lines for research purposes. This process is experimental and the keywords may be updated as the learning algorithm improves. Overview of the lcmsms analyses for eight malignant melanoma cell lines.
It has been administered as a single agent to cutaneous melanoma patients in phase 1 and 2 and 180 patients in expanded access, and is in phase 3 for locally advanced cutaneous melanoma stage iiibiv m1a, nct02288897. The lack of braf, nras, kit, pi3k, and akt mutations reveals gnaq and gna11 uveal melanoma cells to be distinct among melanoma types. Blm and mv3 are highly invasive and metastatic cell lines as evaluated by subcutaneous inoculation into nude mice, 9, 11, 12 whereas a2058 and sk. Semaphorin 3a suppresses tumor growth and metastasis in. Marshall tumor biology laboratory, cancer research uk clinical center, john. Each culture contains genomic mutations in one or more of the following genes according to the sanger cosmic database. Our a375 cells are available with a variety of different reporters, including the human sodium iodide symporter hnis, firefly luciferase fluc, enhanced green. Mouse cell line developed to fasttrack research in braf.
We describe identification of the expression profile of 1,037 mitochondriafocused genes and 84 survivalapoptosis genes in 21 malignant melanoma cell lines and 3 normal melanocyte controls using recently developed hmitchip3 cdna microarrays. By developing cell lines that grow readily in culture, investigators have created a fasttrack research tool that remains applicable to many scientists who use mouse melanoma as a model system. As only few human melanoma cell lines are available that frequently show metastasis in nude mice, the highly metastatic mv3 cell line represents. Our a375 reporter cell lines can be tracked in vivo, making them great tools for studying the mechanisms of tumor growth and metastasis, as well as evaluating the effects of various drugs or therapies in animals. Human melanoma cell lines cls cell lines service part of the cls cell bank table 1. B16 cells are useful models for the study of metastasis and solid tumor formation, and were one of the first effective murine tools for metastasis research. B16 melanoma is a murine tumor cell line used for research as a model for human skin cancers. Description malme3m is a malignant human melanoma cell line that displays fibroblastlike morphology and grows in mixed culture adherentsuspension. In vivo generation of samples for rna sequencing eight weeks old female nod. These unsettling findings highlight the need to authenticate the cell lines that are used in the uveal melanoma field and share this data with other researchers. This cell line has been shown to be dependent upon micropthalmiaassociated transcription factor mitf activity for growth and survival. Established tumor cell lines have been used since the 1970s as an in vitro model for drug discovery. Jci insight tumor cell oxidative metabolism as a barrier.
Plotting the basal metabolism of these cell lines on a 2dimensional plot of oxidative metabolism oxygen consumption rate, ocr versus glycolytic metabolism extracellular acidification rate, ecar revealed that there were distinct metabolic profiles between cell lines. Cancer stem cells cscs have been identified in many cancer types. The data confirm the mutually exclusive nature of gnaq and gna11 mutations in vitro. Three of the melanoma cell lines released an activity that inhibited binding of 125ilabeled pdgf to human foreskin fibroblasts and stimulated 3hthymidine incorporation in such cells. Swetter, md, director of pigmented lesion and cutaneous melanoma clinic. Identification of cancer stem cell subpopulations in head. The human ocular melanoma cell lines, ocm1, ocm1a and om431, were kindly supplied by professor john f. This vector also included an elongation factor alpha promoter ef1a. Kinase inhibitor library screening identifies synergistic. Plotting the basal metabolism of these cell lines on a 2dimensional plot of oxidative metabolism oxygen consumption rate, ocr versus glycolytic metabolism extracellular acidification rate, ecar revealed that there were distinct metabolic profiles between cell lines that have. Excisional biopsy of tumors and subsequent expansion of tumor cell lines has been the traditional tool utilized. A375 human melanoma reporter gene cell lines imanis life. Isolation and characterization of two canine melanoma cell. Tic chromatograms shown in solid black lines are normalized to the maximum value across all.
Semaphorin 3a suppresses tumor growth and metastasis in mice. This article is from journal of translational medicine, volume 9. Brafv600e negatively regulates the akt pathway in melanoma. Genetic and genomic characterization of 462 melanoma. Isolation and characterization of two canine melanoma cell lines. Misidentification of cell lines is not unique to the uveal melanoma research field, as it has been proposed that many cell lines have been misclassified chatterjee, 2007, lacroix, 2008.
Dimethylacrylshikonin induces apoptosis in melanoma cell. Our study demonstrates a new regulatory mechanism between mapk pathway and akt pathway in that brafv600e negatively regulates the akt pathway through interacting with the rictor mtorc2 complex. A375 1 melanoma cell line homo sapiens human caucasian. Herein, we compared the transcriptomes of 42 melanoma cell lines to hundreds of tumours from the cancer. To investigate genetic abnormalities in the context of your gene of interest and other tissues, atcc also offers panels of cell lines organized by genetic alteration. Melanoma is the most dangerous type of skin cancer, and is the leading cause of death from skin disease. Cell reports resource genetic and genomic characterization of 462 melanoma patientderived xenografts, tumor biopsies, and cell lines bradleygarman,1,16 ioannisn. A molecular linkage between the mapk and the lkb1ampk energy sensor. In women, they most commonly occur on the legs, while in men they most commonly occur on the back. Patient derived tumor models and melanoma cell lines. The replicates for all cell lines are ordered as follows.
Investigating the utility of human melanoma cell lines as. B16f10 mouse melanoma cells were cultured in dulbeccos modified eagles medium dmem supplemented. This study identified and characterized cscs in head and neck metastatic malignant melanoma hnmmm to regional lymph nodes using induced pluripotent stem cell ipsc markers. This cell line has been reported to be nonpermissive for human cytomegalovirus hcmv.
Melanoma, also known as malignant melanoma, is a type of cancer that develops from the pigmentproducing cells known as melanocytes. Isolation and culture of melanoma cell lines springerlink. Cells free fulltext identification of cancer stem cell. Melanoma cell melanoma cell line tumor nodule excisional biopsy fine needle aspiration biopsy these keywords were added by machine and not by the authors. Melanoma is not the most common form of skin cancer, but rates are steadily increasing. Genetic and molecular characterization of uveal melanoma. We first employed seahorse analysis to examine various melanoma cell lines that are used in human cancer biology. Our data indicate that melanoma cell lines express discrete stem cell markers.
Plx4032 decreased signaling through the mapk pathway only in cell lines with the brafv600e mutation. Herein, we have analyzed ctla4 expression and ipilimumab reactivity on melanoma cell lines and. Melanoma, also known as malignant melanoma, is a type of cancer that develops from the pigmentcontaining cells known as melanocytes. Total rna samples were extracted from 21 malignant melanoma cell lines and 3 controls and labeled for microarray analysis using hmitchip3. Genetic and molecular characterization of uveal melanoma cell. The transduced cell lines were used in all experiments, authenticated within the last six months using short tandem repeat str profiling, and maintained as previously described 57.
Overall, transcriptional profiles of melanoma cell lines generally resemble that of tumours, with a. Ht144 cells form xenograft tumors when injected into immunocompromised mice. Melanomas typically occur in the skin but may rarely occur in the mouth, intestines or eye uveal melanoma. Immunohistochemical ihc staining performed on 20 hnmmm tissue samples demonstrated expression of ipsc markers. Only a few canine melanoma cell lines have already been reported in the literature. Cell culture protocol for patientderived melanoma tumor cell.
Inhibition of mitochondrial respiration prevents brafmutant. Pdf establishment and characterization of human melanoma cell. Deviating from the culture conditions required for a particular melanoma cell line can result in different phenotypes being expressed. We tested the antitumor effects of a specific inhibitor of raf protein kinases, plx4032rg7204, in melanoma cell lines. Functional features of cancer stem cells in melanoma cell lines. The engagement of ctla4 on primary melanoma cell lines. Differential sensitivity of melanoma cell lines with braf. This chapter summarizes some of the commonly used and better characterized melanoma cell lines, from a total of more than 350 established in this laboratory. The invasive potential of human melanoma cell lines. Hello, im working on melanoma and im searching the best melanoma cell lines easy to culture recapitulating all the steps of melanoma growth. Cells free fulltext upar knockout results in a deep. Human melanoma cell lines of primary and metastatic origin express. We report the mutation status of relevant melanoma genes, expression levels of proteins of interest, and dna fingerprinting of a panel of uveal melanoma cell lines used in the research community. Materials and methods cell lines, reagents and antibodies the murine melanoma cell lines b16f1 and b16f10 and.
Clusters and subclusters of 21 malignant melanoma cell lines revealed by unsupervised hierarchical clustering analysis of 1,037 informative genes from 81 microarray experiments. Functional features of cancer stem cells in melanoma cell. Jan 11, 2012 cell lines from uveal melanoma tumors are an essential tool for these types of analyses. A panel of 4 metastatic melanoma cancer cell lines with varying degrees of genetic complexity. Atcc has developed two melanoma cancer cell panels with varying degrees of genetic complexity, representing both primary and metastatic tumors. The basics of cell culture for patientderived melanoma cell lines share certain similarities, however, cell culture conditions vary typically for each melanoma cell line. Effect of dabrafenib on melanoma cell lines harbouring the.703 1186 414 471 1420 1498 1647 947 166 261 623 1533 171 1329 1105 1319 716 1029 1365 1422 1198 253 1464 1013 884 870 774 1383 498 543 1033 826