scholarly journals Tumor Suppressor Role of hsa-miR-193a-3p and -5p in Cutaneous Melanoma

2020 ◽  
Vol 21 (17) ◽  
pp. 6183
Author(s):  
Beatrice Polini ◽  
Sara Carpi ◽  
Stefano Doccini ◽  
Valentina Citi ◽  
Alma Martelli ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Cutaneous Melanoma ◽  
Luciferase Assay ◽  
Strong Increase ◽  
Over Expression ◽  
Functional Features ◽  
Melanoma Patients ◽  
Melanoma Cell Lines

Background: Remarkable deregulation of several microRNAs (miRNAs) is demonstrated in cutaneous melanoma. hsa-miR-193a-3p is reported to be under-expressed in tissues and in plasma of melanoma patients, but the role of both miR-193a arms in melanoma is not known yet. Methods: After observing the reduced levels of miR-193a arms in plasma exosomes of melanoma patients, the effects of hsa-miR-193a-3p and –5p transfection in cutaneous melanoma cell lines are investigated. Results: In melanoma cell lines A375, 501Mel, and MeWo, the ectopic over-expression of miR-193a arms significantly reduced cell viability as well as the expression of genes involved in proliferation (ERBB2, KRAS, PIK3R3, and MTOR) and apoptosis (MCL1 and NUSAP1). These functional features were accompanied by a significant downregulation of Akt and Erk pathways and a strong increase in the apoptotic process. Since in silico databases revealed TROY, an orphan member of the tumor necrosis receptor family, as a potential direct target of miR-193a-5p, this possibility was investigated using the luciferase assay and excluded by our results. Conclusions: Our results underline a relevant role of miR-193a, both -3p and -5p, as tumor suppressors clarifying the intracellular mechanisms involved and suggesting that their ectopic over-expression could represent a novel treatment for cutaneous melanoma patients.

scholarly journals MicroRNA-603 Promotes Progression of Cutaneous Melanoma by Regulating TBX5

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Xianghua Dong ◽  
Ying Wang ◽  
Yan Qu ◽  
Junru Liu ◽  
Xien Feng ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Cutaneous Melanoma ◽  
Melanoma Cells ◽  
Malignant Progression ◽  
Luciferase Assay ◽  
Luciferase Reporter ◽  
Qrt Pcr ◽  
Gene Assay ◽  
Melanoma Cell Lines

Background. Although studies manifested that microRNA-603 plays a vital role in many cancers, the modulatory mechanism of microRNA-603 in cutaneous melanoma remains unknown. We aimed to investigate the roles of microRNA-603 in cutaneous melanoma cells. Methods. First, microRNA-603 expression in cutaneous melanoma cell lines was detected by qRT-PCR. The mRNA and protein expression levels of TBX5 in cutaneous melanoma cell lines were tested by qRT-PCR and western blot, respectively. In addition, the interaction between microRNA-603 and TBX5 was determined by dual-luciferase reporter gene assay, and their impacts on the growth of cutaneous melanoma cells were detected by cellular function experiments such as MTT, colony formation, and Transwell assays. Results. The expression level of microRNA-603 in human cutaneous melanoma cells was relatively upregulated. Overexpressing microRNA-603 could promote progression of cutaneous melanoma cells, while silencing microRNA-603 expression could suppress the malignant progression of cutaneous melanoma. In addition, TBX5 was lowly expressed in cutaneous melanoma cells. As confirmed by dual-luciferase assay, microRNA-603 could specifically bind to 3 ′ UTR of TBX5 and regulate TBX5. The results of the rescue experiment demonstrated that inhibiting microRNA-603 expression could suppress the proliferation, migration, and invasion of cutaneous melanoma cells, but its suppressive effect could be restored by TBX5. Conclusion. MicroRNA-603 could regulate the expression of TBX5, thus promoting the malignant progression of cutaneous melanoma cells.

scholarly journals Expression of cd44 splice variants in human cutaneous melanoma and melanoma cell lines is related to tumor progression and metastatic potential

1995 ◽  
Vol 64 (3) ◽  
pp. 182-188 ◽  
Author(s):  
Eveliene Manten-Horst ◽  
Erik H. J. Danen ◽  
Lia Smit ◽  
Margriet Snoek ◽  
I. Le Caroline Poole ◽  
...  
Keyword(s):  
Tumor Progression ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Cutaneous Melanoma ◽  
Splice Variants ◽  
Metastatic Potential ◽  
Melanoma Cell Lines

scholarly journals The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas.

1993 ◽  
Vol 178 (2) ◽  
pp. 489-495 ◽  
Author(s):  
V Brichard ◽  
A Van Pel ◽  
T Wölfel ◽  
C Wölfel ◽  
E De Plaen ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Autologous Tumor ◽  
Cytolytic T Lymphocytes ◽  
Pilot Studies ◽  
Tyrosinase Gene ◽  
Melanoma Patients ◽  
Melanoma Cell Lines

Lymphocytes of melanoma patients can be restimulated in vitro with autologous tumor cells to generate antitumor cytolytic T lymphocytes (CTL). Previous reports have indicated that, when such CTL are obtained from HLA-A2 melanoma patients, they often display broad reactivity on A2 melanoma cell lines. Such antitumor CTL clones, which appeared to recognize the same antigen, were isolated from two patients. We report here the cloning of a cDNA that directs the expression of the antigen recognized by these CTL. This cDNA corresponds to the transcript of the tyrosinase gene. The gene was found to be active in all tested melanoma samples and in most melanoma cell lines. Among normal cells, only melanocytes appear to express the gene. The tyrosinase antigen presented by HLA-A2 may therefore constitute a useful target for specific immunotherapy of melanoma. But possible adverse effects of antityrosinase immunization, such as the destruction of normal melanocytes and its consequences, will have to be examined before clinical pilot studies can be undertaken.

scholarly journals Rapid Screening of 4000 Individuals for Germ-line Variations in the BRAF Gene

2006 ◽  
Vol 52 (9) ◽  
pp. 1675-1678 ◽  
Author(s):  
Michael R James ◽  
Troy Dumeni ◽  
Mitchell S Stark ◽  
David L Duffy ◽  
Grant W Montgomery ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Germ Line ◽  
Rapid Screening ◽  
Braf Gene ◽  
Flight Mass Spectrometry ◽  
Melanoma Patients ◽  
Twin Families ◽  
And Control ◽  
Melanoma Cell Lines

Abstract Background: The BRAF gene is frequently somatically altered in malignant melanoma. A majority of variations are at the valine 600 residue leading to a V600E substitution that constitutively activates the kinase. We screened 4000 patient and control DNAs for germ-line variations at the valine 600 residue. Methods: We developed a novel assay by adapting single-base variation assays and software for MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry to screen for all 5 reported variants at codon 600 of the BRAF gene. We screened a case-control collection comprising samples from 1082 melanoma patients and 154 of their unaffected relatives from 1278 families and from 2744 individuals from 659 unselected twin families with no history of melanoma. A panel of 66 melanoma cell lines was used for variation-positive controls. Results: All melanoma cell lines that we had found previously to carry a codon 600 variation were verified in this study. Three of the 4 possible variants (V600E n = 47, V600K n = 2, V600R n = 1) were detected, but no case of V600D was available. No germ-line variants were found in the samples from the 3980 melanoma patients or from the control individuals. Conclusions: This new assay is a high-throughput, automated alternative to standard sequencing and can be used as a rapid initial screen for somatic variants associated with melanoma. Germ-line variants at valine 600 are unlikely to exist and do not contribute to the reported role of the BRAF gene in melanoma predisposition.

Autophagy inhibitors chloroquine and LY294002 enhance temozolomide cytotoxicity on cutaneous melanoma cell lines in vitro

2017 ◽  
Vol 28 (3) ◽  
pp. 307-315 ◽  
Author(s):  
Oxana O. Ryabaya ◽  
Andrey N. Inshakov ◽  
Angelina V. Egorova ◽  
Marina A. Emelyanova ◽  
Tatiana V. Nasedkina ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Cutaneous Melanoma ◽  
Melanoma Cell Lines

scholarly journals Possible role of galectin-9 in cell aggregation and apoptosis of human melanoma cell lines and its clinical significance

2002 ◽  
Vol 99 (6) ◽  
pp. 809-816 ◽  
Author(s):  
Toshiro Kageshita ◽  
Yumiko Kashio ◽  
Akira Yamauchi ◽  
Masako Seki ◽  
Mohammad Jainul Abedin ◽  
...  
Keyword(s):  
Clinical Significance ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Cell Aggregation ◽  
Human Melanoma ◽  
Human Melanoma Cell ◽  
Melanoma Cell Lines

scholarly journals Rho-mediated gene transcription promotes BRAF inhibitor resistance in de-differentiated melanoma cells

2018 ◽  
Author(s):  
SA Misek ◽  
KM Appleton ◽  
TS Dexheimer ◽  
EM Lisabeth ◽  
RS Lo ◽  
...  
Keyword(s):  
Cell Lines ◽  
Gene Transcription ◽  
Melanoma Cell ◽  
Cutaneous Melanoma ◽  
Acquired Resistance ◽  
Rho Kinase ◽  
Braf Inhibitor ◽  
Resistant Cell ◽  
Melanoma Cell Lines

AbstractOver half of cutaneous melanoma tumors have BRAFV600E/Kmutations. Acquired resistance to BRAF inhibitors (BRAFi) remains a major hurdle in attaining durable therapeutic responses. In this study we demonstrate that approximately 50-60% of melanoma cell lines with vemurafenib resistance acquiredin vitroshow activation of RhoA family GTPases. In BRAFi-resistant melanoma cell lines and tumors, activation of RhoA is correlated with decreased expression of melanocyte lineage genes. Using a machine learning approach, we built gene expression-based models to predict drug sensitivity for 265 common anti-cancer compounds. We then projected these signatures onto the collection of TCGA cutaneous melanoma and found that poorly differentiated tumors were predicted to have increased sensitivity to multiple Rho kinase (ROCK) inhibitors. Two transcriptional effectors downstream of Rho, MRTF and YAP1, are activated in the RhoHighBRAFi-resistant cell lines, and resistant cells are more sensitive to inhibition of these transcriptional mechanisms. Taken together, these results support the concept of targeting Rho-regulated gene transcription pathways as a promising therapy approach to restore sensitivity to BRAFi-resistant tumors or as a combination therapy to prevent the onset of drug resistance.

scholarly journals Vemurafenib resistant melanoma cells acquire mesenchymal stem cell-like properties

2019 ◽  
Vol 6 (4) ◽  
pp. 47-57
Author(s):  
A. A. Vartanian ◽  
O. S. Burova ◽  
Kh. S. Vishnyakova ◽  
I. V. Samoylenko ◽  
V. A. Misyurin ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Acquired Resistance ◽  
Melanoma Cells ◽  
Mapk Signaling ◽  
Brafv600e Mutation ◽  
Acquired Drug Resistance ◽  
Melanoma Patients ◽  
Braf Mutant ◽  
Melanoma Cell Lines

Background. Activating mutations in the BRAF gene leads to a constitutive activation of the MAPK signaling. The highly selective BRAFV600E inhibitor, vemurafenib, improves the overall survival of BRAF-mutant melanoma patients. However, despite the excellent results of response rate, the average duration of the response was short and acquired resistance develops in most BRAF mutated melanoma patients within a few months. Objective: to derive melanoma cell lines from surgical species of patients with BRAF mutant melanomas resistant to vemurafenib and to elucidate the mechanisms involved in acquired drug resistance.Materials and methods. Mel Ki and Mel F1702 melanoma cells were obtained from metastases of disseminated melanoma patients with BRAFV600E mutation. 2D tumor cell culture, MTT test, immunicytochemistry, flow cytometry, real-time polimerase chain reaction and osteogenic and adipocytic differentiation were used in the study.Results. We have derived two melanoma cell lines Mel Ki and Mel F1702 from tumor samples of patients with BRAFV600E mutation resistant to vemurafenib. These cells were homogenous and had fibroblastic morphology. The IC50 values for Mel Ki and Mel F1702 were 4.7 and 6.3 μM, respectively. The expression of cancer-testis antigens was not detected in both types of cells suggesting the stemness of Mel Ki and Mel F1702 melanoma cells. The immunophenotypic profile of the vemurafenib resistsant melanoma cells showed the expression of typical mesenchymal stem cells markers such as CD90, CD105 and CD44. In addition, we found that the melanoma cell lines derived from tumor resistant to vemurafenib differentiated into osteoblastand adipocyte-like cells. Conclusion. In this study we are offering an experimental evidence of the phenotypic transition of the vemurafenib-resistant melanoma cells into mesenchymal stem-like cells.

scholarly journals LncRNA MEG3 promotes melanoma growth, metastasis and formation through modulating miR-21/E-cadherin axis

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Liangcai Wu ◽  
Lifei Zhu ◽  
Yanchang Li ◽  
Zhixin Zheng ◽  
Xi Lin ◽  
...  
Keyword(s):  
Tumor Growth ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Tumor Metastasis ◽  
Luciferase Reporter ◽  
Rt Pcr ◽  
Kaplan Meier ◽  
Melanoma Cell Lines ◽  
E Cadherin

Abstract Background Melanoma is the most aggressive type of skin cancer with high mortality rate and poor prognosis. lncRNA MEG3, a tumor suppressor, is closely related to the development of various cancers. However, the role of lncRNA MEG3 in melanoma has seldom been studied. Methods RT-PCR was used to examine the expressions of lncRNA MEG3 and E-cadherin in melanoma patients and cell lines. Then, the biological functions of lncRNA MEG3 and E-cadherin were demonstrated by transfecting lncRNA MEG3-siRNA, lncRNA MEG3-overexpression, E-cadherin-siRNA and E-cadherin-overexpression plasmids in melanoma cell lines. Moreover, CCK8 assay and colony formation assay were utilized to assess the cell proliferation; Transwell assay was performed to evaluate the cell invasive ability; and tumor xenografts in nude mice were applied to test the tumor generation. Additionally, the target interactions among lncRNA MEG3, miR-21 and E-cadherin were determined by dual luciferase reporter assay. Finally, RT-PCR and WB were further conducted to verify the regulatory roles among lncRNA MEG3, miR-21 and E-cadherin. Results The clinical data showed that lncRNA MEG3 and E-cadherin expressions were both declined in carcinoma tissues as compared with their para-carcinoma tissues. Moreover, lncRNA MEG3 and E-cadherin expressions in B16 cells were also higher than those in A375 and A2058 cells. Subsequently, based on the differently expressed lncRNA MEG3 and E-cadherin in these human melanoma cell lines, we chose B16, A375 and A2058 cells for the following experiments. The results demonstrated that lncRNA MEG3 could suppress the tumor growth, tumor metastasis and formation; and meanwhile E-cadherin had the same effects on tumor growth, tumor metastasis and formation. Furthermore, the analysis of Kaplan–Meier curves also confirmed that there was a positive correlation between lncRNA MEG3 and E-cadherin. Ultimately, dual luciferase assays were further used to verify that lncRNA MEG3 could directly target miR-21 which could directly target E-cadherin in turn. Additionally, the data of RT-PCR and WB revealed that knockdown of lncRNA MEG3 in B16 cells inhibited miR-21 expression and promoted E-cadherin expression, but overexpression of lncRNA MEG3 in A375 and A2058 cells presented completely opposite results. Conclusion Our findings indicated that lncRNA MEG3 might inhibit the tumor growth, tumor metastasis and formation of melanoma by modulating miR-21/E-cadherin axis.

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