primary a375 78 sk mel 28  (ATCC)

Journal: Oncology Letters

Article Title: Role of ELP6 in tumour progression and impact on ERK1/2 signalling pathway inhibitors in skin cutaneous melanoma

doi: 10.3892/ol.2025.14996

Figure Lengend Snippet: ELP6 is essential for driving SKCM progression. (A) ELP6 knockdown efficiencies were assessed in 293T cells using two independent shRNAs, shELP6-2 and shELP6-3. The viability of shCtrl, (B) shELP6-2 and (C) shELP6-3 cells was assessed at 0 and 24 h. (D) ELP6 knockdown efficiency using shELP6 was assessed in A375 cells. (E) The viability of shCtrl and shELP6 A375 cells was assessed at 0, 24 and 48 h. The relative mRNA expression levels of Ki67 (F and G) and PCNA (H and I) were evaluated in shCtrl, shELP6-2 and shELP6-3 293T cells, as well as in shELP6 A375 cells. The levels of PCNA mRNA expression levels were evaluated in (J) shELP6-2 and (K) shELP6-3 cells after transfection with either the pEGFP-C2 or pEGFP-ELP6 plasmid. (L) Cell cycle distribution was analysed using flow cytometry, and the percentage of cells in each phase of the cell cycle was calculated. (M) shCtrl and (N) shELP6 A375 cells were exposed to 10% serum for 24 h, followed by staining with propidium iodide. *P<0.05; **P <0.01; ***P<0.001. ELP6, elongator acetyltransferase complex subunit 6; SKCM, skin cutaneous melanoma; sh, short hairpin; m, messenger; Ctrl, control; PCNA, proliferating cell nuclear antigen.

Article Snippet: The 293T, SK-MEL-2 and A375 cell lines were obtained from Procell Life Science & Technology Co., Ltd., and were authenticated using STR analysis.

Techniques: Knockdown, Expressing, Transfection, Plasmid Preparation, Flow Cytometry, Staining, Control

Journal: Oncology Letters

Article Title: Role of ELP6 in tumour progression and impact on ERK1/2 signalling pathway inhibitors in skin cutaneous melanoma

doi: 10.3892/ol.2025.14996

Figure Lengend Snippet: p42 MAPK serves a key role in mediating the proliferative effects induced by ELP6 in SKCM. Western blot analysis in shCtrl, (A) shELP6-3 293T and (B) shELP6 A375 cells of p42 MAPK or pi-p42 MAPK (Thr202/Tyr204) expression levels. (C and D) Quantification of p42 MAPK or pi-p42 MAPK (Thr202/Tyr204) protein expression levels. mRNA expression levels of p44 MAPK and p42 MAPK, normalized to the GAPDH, in the shCtrl, (E) shELP6-2, shELP6-3 293T and (F) sh-ELP6 A375 cells. Western blot analysis of ELP6-GFP, tubulin and p42 MAPK in (G) shELP6-3 293T and (H) shELP6 A375 cells transfected with either the pEGFP-C2 or pEGFP-ELP6 plasmids, with quantification of p42 MAPK protein expression levels. pcDNA3.10V5-HisB or p42 MAPK-pcDNA3.10V5-HisB transfected into shCtrl, (I) shELP6-3 293T and (J) shELP6 A375 cells were subject to western blot analysis of p42 MAPK expression levels and (K and L) cell viability assays under the same treatment conditions. *P<0.05; **P<0.01; ***P<0.001. ELP6, elongator acetyltransferase complex subunit 6; ns, not significant; pi, phosphorylated; SKCM, skin cutaneous melanoma; sh, short hairpin; Ctrl, control.

Article Snippet: The 293T, SK-MEL-2 and A375 cell lines were obtained from Procell Life Science & Technology Co., Ltd., and were authenticated using STR analysis.

Techniques: Western Blot, Expressing, Transfection, Control

Journal: Oncology Letters

Article Title: Role of ELP6 in tumour progression and impact on ERK1/2 signalling pathway inhibitors in skin cutaneous melanoma

doi: 10.3892/ol.2025.14996

Figure Lengend Snippet: ELP6 stimulates p42 MAPK at the post-transcriptional level. (A) shCtrl and shELP6-3 293T cell lines were treated with CHX followed by western blotting for p42 MAPK and tubulin, quantified in (B). (C) shCtrl and shELP6 A375 cells lines were similarly treated, followed by western blotting for p42 MAPK and GAPDH, quantified in (D). The (E) 293T and (F) A375 cell lines were treated with siT4 or siNC and mRNA expression levels of LysRS, normalized to the GAPDH, were determined to detect knockdown efficiencies. p42 MAPK protein expression levels, normalized to tubulin, were determined in the (G) 293T and (H) A375 cell lines treated with siT4 or siNC. *P<0.05. ns, not significant; NC, negative control; t, transfer; sh, short hairpin; si, silencing; LysRS, lysyl tRNA synthetase; ELP6, elongator acetyltransferase complex subunit 6; SKCM, skin cutaneous melanoma; Ctrl, control; CHX, cycloheximide; siT4, LysRS siRNA.

Article Snippet: The 293T, SK-MEL-2 and A375 cell lines were obtained from Procell Life Science & Technology Co., Ltd., and were authenticated using STR analysis.

Techniques: Western Blot, Expressing, Knockdown, Negative Control, Control

Journal: Oncology Letters

Article Title: Role of ELP6 in tumour progression and impact on ERK1/2 signalling pathway inhibitors in skin cutaneous melanoma

doi: 10.3892/ol.2025.14996

Figure Lengend Snippet: ELP6 modulates drug sensitivity in melanoma cell lines. Box plots of the predicted clinical sensitivity of (A) AZ628, (B) Sorafenib, (C) NVP.BEZ235 and (D) VX702 in patients with SKCM categorized into ELP6 high (red) or ELP6 low (blue) groups from the The Cancer Atlas Genome database (n=468). (E) shCtrl, shELP6-2 and shELP6-3 293T cells were treated with U0126 for 0 and 48 h, and CCK-8 assays were performed to measure cell viability. shCtrl and shELP6 A375 cells were treated with U0126 for (F) 0, 24 and (G) 48 h, followed by a CCK-8 assay to assess cell viability. (H and I) shELP6-3 293T and shELP6 A375 cell lines were transfected with either the pEGFP-C2 or pEGFP- ELP6 plasmid, and treated U0126 for 48 h, followed by a CCK8 assay to evaluate cell viability. shCtrl (J) −293T and (K) -A375 cells were treated with U0126 for 24 h, ELP6 mRNA levels expression levels, normalized to GAPDH, were assessed. (L) Schematic illustrating the modulation of ERK1/2 by ELP6. Data are presented as mean ± SEM. Error bars represent the SEM and each point in the graph corresponds to an individual sample. *P<0.05; **P<0.01; ***P<0.001. ns, not significant; ELP6, elongator acetyltransferase complex subunit 6; SKCM, skin cutaneous melanoma; Ctrl, control; CCK-8, cell counting kit-8; Sh, short hairpin; SEM, standard error of the mean.

Article Snippet: The 293T, SK-MEL-2 and A375 cell lines were obtained from Procell Life Science & Technology Co., Ltd., and were authenticated using STR analysis.

Techniques: CCK-8 Assay, Transfection, Plasmid Preparation, Expressing, Control, Cell Counting

Journal: Journal of Inflammation Research

Article Title: Envoplakin Inhibits Macrophage Polarization by Altering the Inflammatory Tumor Microenvironment of Melanoma Through the RAS / ERK Signaling Pathway

doi: 10.2147/JIR.S447934

Figure Lengend Snippet: EVPL regulates the malignant progression of melanoma and the inflammatory tumor microenvironment of melanoma. ( A ) Expression changes of EVPL were detected by Western blot, n=3. ( B ) CCK-8 assay was used to detect the viability of A375 and A2058 cells, n = 3. ( C and D ) Colony formation assay was used to detect the proliferation of A375 and A2058 cells, n=3. ( E and F ) Trans-well invasion assay was used to detect the invasion of A375 and A2058 cells, n = 3. ( G and H ) Wound-healing assay was used to detect the migration of A375 and A2058 cells, n=3. ( I and J ) Flow cytometry was used to detect the apoptosis rate of A375 and A2058 cells, n=3. ( K and L ) ELISA was used to detect the expression of tumor immune-related chemokines and inflammatory factors in the cell supernatant, n=3. The data in the figures represent the mean±SD. * Represents P < 0.05, ** Represents P <0.01, *** Represents P <0.001.

Article Snippet: A375, A2058, A875, and Sk-mel-28 (melanoma cells) were purchased from Procell Life Science & Technology Co., Ltd. HaCaT (normal cells) were procured from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Expressing, Western Blot, CCK-8 Assay, Colony Assay, Invasion Assay, Wound Healing Assay, Migration, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: JCI Insight

Article Title: Differential histone acetylation and super-enhancer regulation underlie melanoma cell dedifferentiation

doi: 10.1172/jci.insight.166611

Figure Lengend Snippet: RNA-Seq of Epgn1 cell lines ( n = 3; WM35, YUPEET, WM983A) and Epgn3 cell lines ( n = 2; WM1552C, YUCHIME). The top bar indicates cellular subtypes (Epgn1 [red] and Epgn3 [light blue]) as characterized by the 122-epigenetic signature . Each row of the heatmap indicates a differentially expressed gene, and each column represents a BRAFV600 mutant cell line ( n = 5; each in triplicate). Differentially expressed genes are significant if q < 0.05 by Benjamini-Hochberg procedure and a linear fold-change ± 1.5. The heatmaps are color-coded on the basis of Z scores. ( A ) Supervised hierarchical clustering of 122 epigenetic genes and identification of Epgn1 and Epgn3 groups. ( B ) RNA-Seq analysis identifies increased expression of proliferative and differentiation genes ( MLANA, TYR, DCT, MITF ) in the Epgn1 group and increased expression of invasive genes ( WNT5A, ITGA2, ZEB1, EGFR, ITGA3, PDGFC, NRP, AXL ) in the Epgn3 group using the Hoek proliferative and invasive gene signature . ( C ) Pigmentation pathway genes ( MITF, MLANA, PMEL, TYR, DCT, TYRP1 ) were uniformly upregulated in Epgn1 cells. ( D and E ) Differential expression of genes involved in epithelial mesenchymal transition (EMT) and integrin signaling. ( F ) Heatmap of 51 significantly differentially expressed proteins (after multitesting correction at FDR 5%) determined by reverse phase protein array (RPPA) coincides with transcriptional data. ( G ) GSEA pathway analysis. Pathways are abbreviated as follows: K, KEGG; R, Reactome; H, Hallmark; P, Pathway Interaction Database. ( H ) RNA-Seq data set of TCGA melanomas ( n = 473). ssGSEA analysis depicting correlation between the upregulated Epng1 gene signature with the proliferative genes ( P = 1.069 × 10 –6 ; OR, 2.61), and the upregulated Epgn3 signature with the invasive genes ( P = 8.595 × 10 –6 ; OR, 2.46).

Article Snippet: Metastatic cell lines SKmel2, SKmel5, and A375 were obtained from ATCC.

Techniques: RNA Sequencing, Mutagenesis, Expressing, Quantitative Proteomics, Protein Array