Journal: Cell adhesion & migration

Article Title: Expression of kindlin-3 in melanoma cells impedes cell migration and metastasis.

doi: 10.1080/19336918.2016.1243645

Figure Lengend Snippet: Figure 6. Analyses of kindlin-3 expression in M10 human melanoma cell line. (A) qRT-PCR analyses of kindlin-1, ¡2 and ¡3 mRNA expression levels in M10 cells and the human chronic myeloid leukemia cell line K562. Values are mean § SD of technical triplicates. A representative experiment from 3 independent experiments is shown. (B) qRT-PCR analyses of kindlin-3 expression in M10, K562 and the human fibroblast cell line 293T. In (A) and (B), GAPDH and actin were used as internal controls. (C) Immunoblot analyses of kindlin-3 protein expression in M10 cells, K562, 293T and the human breast cancer cell line MDA-MB-231 using the anti-kindlin-3 mAb clone 9. Actin serves as loading control. (D) Immunoblot analyses using the anti-kindlin-3 mAb 3D6.

Article Snippet: Human melanoma cell line M10 was purchased from American Type Culture Collection (Manassas, VA) and cultured in RPMI1640 fullmedium containing supplements aforementioned.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Cell adhesion & migration

Article Title: Expression of kindlin-3 in melanoma cells impedes cell migration and metastasis.

doi: 10.1080/19336918.2016.1243645

Figure Lengend Snippet: Figure 7. Analyses of the migratory properties of M10 cells transfected with GFP-kindlin-3. (A) Flow cytometry analyses and sorting of cells transiently transfected with GFP or GFP-kindlin-3 expression plasmid. (B) Sorted cells were subjected to 2D-random migration assays on fibronectin-coated (2 mg/cm2) coverslip-bottom culture dishes. Twenty cells were tracked and migratory profiles were plotted. Representative data from at least 2 independent experiments are shown. X and Y axes are relabeled with larger fonts for clarity.

Article Snippet: Human melanoma cell line M10 was purchased from American Type Culture Collection (Manassas, VA) and cultured in RPMI1640 fullmedium containing supplements aforementioned.

Techniques: Transfection, Flow Cytometry, Expressing, Plasmid Preparation, Migration

Journal: Molecular Medicine Reports

Article Title: Rsf-1 regulates malignant melanoma cell viability and chemoresistance via NF-κB/Bcl-2 signaling

doi: 10.3892/mmr.2019.10610

Figure Lengend Snippet: Rsf-1 expression in melanoma cell lines and Rsf-1 knockdown efficiency. (A) Western blotting and RT-qPCR analysis revealed the endogenous expression levels of Rsf-1 in three melanoma cell lines (MV3, M14 and A375). (B) Western blotting and RT-qPCR analysis demonstrated that Rsf-1 siRNA transfection significantly decreased Rsf-1 expression levels in MV3 and A375 cells, while Rsf-1 plasmid transfection upregulated the protein and mRNA expression of Rsf-1 in M14 cells. Data were presented as the mean ± standard deviation of at least three experiments. *P<0.05 vs. control. Rsf-1, remodeling and spacing factor 1; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; siRNA, small interfering RNA.

Article Snippet: The targeting sequences were as follows: Rsf-1 siRNA, 5′-GGAAAGACAUCUCUACUAU-3′; and control siRNA, 5′-GCGCGATAGCGCGAATATA-3′. pCMV6-Rsf-1 and control empty plasmids were purchased from OriGene Technologies, Inc. (Rockville, MD, USA), and M14 cells were transfected with 1 µg plasmid using Lipofectamine 3000 according to the manufacturer's protocols.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Transfection, Plasmid Preparation, Standard Deviation, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: Rsf-1 regulates malignant melanoma cell viability and chemoresistance via NF-κB/Bcl-2 signaling

doi: 10.3892/mmr.2019.10610

Figure Lengend Snippet: Rsf-1 regulates melanoma cell viability and invasion. (A) An MTT assay (96-well plate) revealed that Rsf-1 depletion decreased the viability of MV3 and A375 cells; conversely, Rsf-1 overexpression increased M14 cell viability. (B) A colony formation assay (culture dish diameter, 6 cm) demonstrated that the colony number was reduced in MV3 and A375 cells transfected with Rsf-1 siRNA, while Rsf-1 overexpression promoted colony formation ability in M14 cells. (C) A Transwell invasion assay (24-well plate) revealed that the number of invading cells decreased following Rsf-1 depletion in MV3 and A375, and increased following Rsf-1 overexpression in M14 cells. Data were presented as the mean ± standard deviation of at least three experiments. *P<0.05 vs. control. Magnification, ×200. Rsf-1, remodeling and spacing factor 1; siRNA, small interfering RNA.

Article Snippet: The targeting sequences were as follows: Rsf-1 siRNA, 5′-GGAAAGACAUCUCUACUAU-3′; and control siRNA, 5′-GCGCGATAGCGCGAATATA-3′. pCMV6-Rsf-1 and control empty plasmids were purchased from OriGene Technologies, Inc. (Rockville, MD, USA), and M14 cells were transfected with 1 µg plasmid using Lipofectamine 3000 according to the manufacturer's protocols.

Techniques: MTT Assay, Over Expression, Colony Assay, Transfection, Transwell Invasion Assay, Standard Deviation, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: Rsf-1 regulates malignant melanoma cell viability and chemoresistance via NF-κB/Bcl-2 signaling

doi: 10.3892/mmr.2019.10610

Figure Lengend Snippet: Rsf-1 regulates cell cycle progression of melanoma and expression of MMP2, cyclin E and p-IκB. (A) Cell cycle analysis revealed that Rsf-1 depletion increased the percentage of G1 phase cells and decreased that of S phase cells in MV3 and A375 cell groups; Rsf-1 overexpression in M14 cells exhibited opposing effects. (B) Western blotting demonstrated that Rsf-1 depletion decreased the levels of MMP2, cyclin E and p-IκB in MV3 and A375 cell lines. Rsf-1 overexpression upregulated expression of MMP2, cyclin E and p-IκB in M14 cells. Data were presented as the mean ± standard deviation of at least three experiments. *P<0.05 vs. control. MMP2, matrix metalloproteinase-2; p, phosphorylated; Rsf-1, remodeling and spacing factor 1; siRNA, small interfering RNA.

Article Snippet: The targeting sequences were as follows: Rsf-1 siRNA, 5′-GGAAAGACAUCUCUACUAU-3′; and control siRNA, 5′-GCGCGATAGCGCGAATATA-3′. pCMV6-Rsf-1 and control empty plasmids were purchased from OriGene Technologies, Inc. (Rockville, MD, USA), and M14 cells were transfected with 1 µg plasmid using Lipofectamine 3000 according to the manufacturer's protocols.

Techniques: Expressing, Cell Cycle Assay, Over Expression, Western Blot, Standard Deviation, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: Rsf-1 regulates malignant melanoma cell viability and chemoresistance via NF-κB/Bcl-2 signaling

doi: 10.3892/mmr.2019.10610

Figure Lengend Snippet: Rsf-1 regulates chemoresistance and the MMP of melanoma cells. (A) An MTT assay revealed that cell viability was decreased following Rsf-1 depletion in MV3 and A375 cells treated with cisplatin. Rsf-1 overexpression promoted cell viability in M14 cells treated with cisplatin. (B) Annexin V/propidium iodide analysis revealed that the percentage of apoptotic cells was significantly increased in Rsf-1-depleted MV3 and A375 cells compared with controls. Rsf-1 overexpression downregulated cisplatin-induced apoptosis in M14 cells. (C) Rsf-1 overexpression reduced MMP depolarization in M14 cells, while Rsf-1 depletion increased depolarization in MV3 and A375 cells treated with cisplatin. Data were presented as the mean ± standard deviation of at least three experiments. *P<0.05 vs. control. FITC, fluorescein isothiocyanate; MMP, mitochondrial membrane potential, Rsf-1, remodeling and spacing factor 1; siRNA, small interfering RNA.

Article Snippet: The targeting sequences were as follows: Rsf-1 siRNA, 5′-GGAAAGACAUCUCUACUAU-3′; and control siRNA, 5′-GCGCGATAGCGCGAATATA-3′. pCMV6-Rsf-1 and control empty plasmids were purchased from OriGene Technologies, Inc. (Rockville, MD, USA), and M14 cells were transfected with 1 µg plasmid using Lipofectamine 3000 according to the manufacturer's protocols.

Techniques: MTT Assay, Over Expression, Standard Deviation, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: Rsf-1 regulates malignant melanoma cell viability and chemoresistance via NF-κB/Bcl-2 signaling

doi: 10.3892/mmr.2019.10610

Figure Lengend Snippet: Rsf-1 regulates Bcl-2 expression via NF-κB signaling. (A) Western blotting revealed that Bax expression levels increased, whereas cIAP1, cIAP2 and Bcl-2 expression decreased significantly following Rsf-1 depletion in MV3 and A375 cells. Rsf-1 overexpression in M14 cells exhibited opposing effects. (B) NF-κB inhibition significantly downregulated p-IκB and NF-κB p65 protein levels in M14 cells. NF-κB inhibition also eradicated the effects of Rsf-1 overexpression on Bcl-2 upregulation. Total IκB expression was markedly altered. Data were presented as the mean ± standard deviation of at least three experiments. *P<0.05 vs. control. Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma 2; cIAP1, cellular inhibitor of apoptosis protein 1; NF-κB, nuclear factor κ-light-chain-enhancer of activated B cells; p, phosphorylated; Rsf-1, remodeling and spacing factor 1; siRNA, small interfering RNA.

Article Snippet: The targeting sequences were as follows: Rsf-1 siRNA, 5′-GGAAAGACAUCUCUACUAU-3′; and control siRNA, 5′-GCGCGATAGCGCGAATATA-3′. pCMV6-Rsf-1 and control empty plasmids were purchased from OriGene Technologies, Inc. (Rockville, MD, USA), and M14 cells were transfected with 1 µg plasmid using Lipofectamine 3000 according to the manufacturer's protocols.

Techniques: Expressing, Western Blot, Over Expression, Inhibition, Standard Deviation, Small Interfering RNA

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Synthesis, in silico modelling, and in vitro biological evaluation of substituted pyrazole derivatives as potential anti-skin cancer, anti-tyrosinase, and antioxidant agents

doi: 10.1080/14756366.2023.2205042

Figure Lengend Snippet: Cytotoxicity of pyrazole (celecoxib analogs), isoxazole, pyrazolone, and positive control compounds P1 – P25 (structures, Figure 3 ) against cells of human cutaneous melanoma and non-melanoma skin cancer lines relative to standard control noncancerous immortalised HaCaT cells.

Article Snippet: Human-derived GFP-expressing melanoma A375 and epidermoid carcinoma A431 cell lines were purchased from Angio-Proteomie (Boston, MA).

Techniques: Positive Control, Control