Journal: PLoS ONE

Article Title: Deregulation of the MiR-193b-KRAS Axis Contributes to Impaired Cell Growth in Pancreatic Cancer

doi: 10.1371/journal.pone.0125515

Figure Lengend Snippet: (A) The TargetScan online database ( http://www.targetscan.org ) predicted KRAS as an miR-193b target. The site between 1074 and 1080 (boxed region), one of the two predicted miR-193b binding sites in the 3′-UTR of KRAS, is shown, highlighting the evolutionarily conservation among humans (hsa), the chimpanzees (Ptr), rhesus monkey (Mml), bushbaby (Oga), and treeshrew (Tbe). (B) The two miR-193b binding sites at positions 303 to 309 and 1074–1080 in the KRAS 3′-UTR, identified using the TargetScan online database ( http://www.targetscan.org ), are shown. Seed sequences were mutated as shown in the boxed regions. The wild-type or mutant constructs were inserted into the pGL3 vector directly downstream of the luciferase gene. (C) 293A cells were co-transfected with miR-193b or scrambled oligonucleotide, wild-type or mutant firefly luciferase constructs of the KRAS 3′-UTR segment containing miR-193b binding sites, and Renilla luciferase (endogenous control). Luciferase activity was measured 24 hours after transfection using the Dual-Luciferase Reporter Assay Systems. Renilla-normalized luciferase activity is expressed relative to that obtained for the scrambled oligonucleotide under each condition. WT, wild-type; Mut, mutant. (D) KRAS and CCND1 protein levels were assessed by Western blotting 48 hours after transfection. (E) The protein levels of p-ERK and p-Akt were evaluated by Western blotting 48 hours after miR-193b transfection.

Article Snippet: In PDAC cell lines, we found that miR-193b expression is higher in BxPC-3 cells bearing wild-type KRAS compared to the AsPC-1, MIA PaCa-2 and PANC-1 cells, which harbor mutant KRAS (mutation statuses were obtained from www.atcc.org and www.sanger.ac.uk ).

Techniques: Binding Assay, Mutagenesis, Construct, Plasmid Preparation, Luciferase, Transfection, Control, Activity Assay, Reporter Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

doi: 10.3390/ijms23010035

Figure Lengend Snippet: Effect of DOX on cell viability of A375 and MNT-1 cells. Cells were exposed to different concentrations of DOX for 24, 48, and 72 h, and cell viability was determined using MTT assay. Data shown are mean values ± standard deviation of three independent experiments with four technical replicates each. *—indicates statistical significance in comparison to the respective control ( p < 0.05).

Article Snippet: Human melanoma cell line A375 was purchased from the European Collection of Authenticated Cell Cultures (ECACC) and supplied by Sigma-Aldrich (Madrid, Spain) and the MNT-1 melanoma cell line was kindly provided by Dr. Manuela Gaspar (iMed.ULisboa, Lisbon, Portugal).

Techniques: MTT Assay, Standard Deviation, Comparison, Control

Journal: International Journal of Molecular Sciences

Article Title: Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

doi: 10.3390/ijms23010035

Figure Lengend Snippet: Inhibitory concentrations (ICs) obtained for 24, 48, and 72 h DOX exposure. Values are expressed in μM.

Article Snippet: Human melanoma cell line A375 was purchased from the European Collection of Authenticated Cell Cultures (ECACC) and supplied by Sigma-Aldrich (Madrid, Spain) and the MNT-1 melanoma cell line was kindly provided by Dr. Manuela Gaspar (iMed.ULisboa, Lisbon, Portugal).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

doi: 10.3390/ijms23010035

Figure Lengend Snippet: Effect of hyperthermia plus DOX on cell viability of A375 and MNT-1 cells. Cells were exposed to 43 °C for 30, 60, or 120 min, plus 0.012 μM or 0.043 μM and 0.68 μM or 1.38 μM during 24 h; 0.0056 μM or 0.0125 μM and 0.0066 μM or 0.0179 μM during 48 h; and 0.0012 μM or 0.0026 μM and 0.0042 μM or 0.0098 μM during 72 h; in cases of A375 or MNT-1, respectively. DMSO concentrations correspond to the equivalent percentage present in IC 20 of each cell line and time exposure. DOX concentrations correspond to the calculated IC 10 and IC 20 for each time exposure and for each cell line. Cell viability was determined using MTT assay. Data are shown as mean ± standard deviation of two independent experiments with four technical replicates each. *—indicates statistical significance in comparison to the control 37 °C; α indicates statistical significance in comparison to the respective control of each condition at 37 °C; and β indicates statistical significance of the combined treatment in comparison to hyperthermia alone ( p < 0.05).

Article Snippet: Human melanoma cell line A375 was purchased from the European Collection of Authenticated Cell Cultures (ECACC) and supplied by Sigma-Aldrich (Madrid, Spain) and the MNT-1 melanoma cell line was kindly provided by Dr. Manuela Gaspar (iMed.ULisboa, Lisbon, Portugal).

Techniques: MTT Assay, Standard Deviation, Comparison, Control

Journal: International Journal of Molecular Sciences

Article Title: Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

doi: 10.3390/ijms23010035

Figure Lengend Snippet: Effect of hyperthermia plus DOX on morphology of A375 and MNT-1 cells. Cells were exposed to 43 °C for 30 min and 0.0125 μM or 0.0179 μM of DOX, in case of A375 or MNT-1 cells, respectively. ( A )—A375 cells; ( B )—MNT-1 cells.

Article Snippet: Human melanoma cell line A375 was purchased from the European Collection of Authenticated Cell Cultures (ECACC) and supplied by Sigma-Aldrich (Madrid, Spain) and the MNT-1 melanoma cell line was kindly provided by Dr. Manuela Gaspar (iMed.ULisboa, Lisbon, Portugal).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

doi: 10.3390/ijms23010035

Figure Lengend Snippet: Effects of hyperthermia combined with DOX on cell cycle distribution. Cells were exposed to 43 °C for 30 min and 0.0125 μM or 0.0179 μM of DOX, in case of A375 or MNT-1 cells, respectively. ( A ) Cell cycle distribution (%) in A375 and MNT-1 cells; ( B ) histograms representative of cell distribution of A375 and MNT-1 cells. Data shown are mean values ± standard deviation of two independent experiments with two technical replicates each and each replicate with at least 5000 events. *—indicates statistical significance in comparison to the control 37 °C; α indicates statistical significance in comparison to the respective control of each condition at 37 °C; and β indicates statistical significance of the combined treatment in comparison to hyperthermia alone ( p < 0.05).

Article Snippet: Human melanoma cell line A375 was purchased from the European Collection of Authenticated Cell Cultures (ECACC) and supplied by Sigma-Aldrich (Madrid, Spain) and the MNT-1 melanoma cell line was kindly provided by Dr. Manuela Gaspar (iMed.ULisboa, Lisbon, Portugal).

Techniques: Standard Deviation, Comparison, Control

Journal: International Journal of Molecular Sciences

Article Title: Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

doi: 10.3390/ijms23010035

Figure Lengend Snippet: Effects of hyperthermia combined with DOX on production of intracellular ROS. Cells were exposed to 43 °C for 30 min and 0.0125 μM or 0.0179 μM of DOX for 48 h, in case of A375 or MNT-1 cells, respectively. ( A ) Relative abundance of intracellular ROS of A375 and MNT-1 cells; ( B ) histograms representative of abundance of intracellular ROS of A375 and MNT-1 cells. Data shown are mean values ± standard deviation of two independent experiments with two technical replicates each and each replicate with at least 5000 events. *—indicates statistical significance in comparison to the control 37 °C; α indicates statistical significance in comparison to the respective control of each condition at 37 °C; and β indicates statistical significance of the combined treatment in comparison to hyperthermia alone ( p < 0.05).

Article Snippet: Human melanoma cell line A375 was purchased from the European Collection of Authenticated Cell Cultures (ECACC) and supplied by Sigma-Aldrich (Madrid, Spain) and the MNT-1 melanoma cell line was kindly provided by Dr. Manuela Gaspar (iMed.ULisboa, Lisbon, Portugal).

Techniques: Standard Deviation, Comparison, Control

Journal: International Journal of Molecular Sciences

Article Title: Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

doi: 10.3390/ijms23010035

Figure Lengend Snippet: Effects of hyperthermia in combination with DOX on apoptotic profile. Both cell lines were exposed to 43 °C for 30 min and A375 cells were treated with 0.0125 μM and MNT-1 cells with 0.0179 μM of DOX for 48 h. ( A ) Percentage of apoptotic cells after treatment in populations corresponding to viable and non-apoptotic, early and late apoptotic A375 and MNT-1 cells; ( B ) histograms representative of Annexin V-FITC. Data shown are mean values ± standard deviation of two independent experiments with two technical replicates each and each replicate with at least 5000 events. *— indicates statistical significance in comparison to the control 37 °C; α indicates statistical significance in comparison to the respective control of each condition at 37 °C; and β indicates statistical significance of the combined treatment in comparison to hyperthermia alone ( p < 0.05).

Article Snippet: Human melanoma cell line A375 was purchased from the European Collection of Authenticated Cell Cultures (ECACC) and supplied by Sigma-Aldrich (Madrid, Spain) and the MNT-1 melanoma cell line was kindly provided by Dr. Manuela Gaspar (iMed.ULisboa, Lisbon, Portugal).

Techniques: Standard Deviation, Comparison, Control

Journal: Experimental and Therapeutic Medicine

Article Title: The biological effect and mechanism of the Wnt/β-catenin signaling pathway on malignant melanoma A375 cells

doi: 10.3892/etm.2018.6413

Figure Lengend Snippet: Cell morphology observed using an inverted microscope (magnification, ×200). A375-RNAi cells and A375-negative cells exhibited a strong green fluorescence; wheras A375 cells did not. siRNA, short interfering RNA.

Article Snippet: Cell culture and transfection The human MM A375 cell line was purchased from the Biovector Science Lab, Inc. (Beijing, China).

Techniques: Inverted Microscopy, Fluorescence, Small Interfering RNA

Journal: Experimental and Therapeutic Medicine

Article Title: The biological effect and mechanism of the Wnt/β-catenin signaling pathway on malignant melanoma A375 cells

doi: 10.3892/etm.2018.6413

Figure Lengend Snippet: Cellular expression influence of lentiviral vector-mediated siRNA on the expression of β-catenin and β-actin in A375 cells. siRNA, small interfering RNA.

Article Snippet: Cell culture and transfection The human MM A375 cell line was purchased from the Biovector Science Lab, Inc. (Beijing, China).

Techniques: Expressing, Plasmid Preparation, Small Interfering RNA

Journal: Experimental and Therapeutic Medicine

Article Title: The biological effect and mechanism of the Wnt/β-catenin signaling pathway on malignant melanoma A375 cells

doi: 10.3892/etm.2018.6413

Figure Lengend Snippet: Influence of lentiviral vector-mediated siRNA on A375 cell proliferation.

Article Snippet: Cell culture and transfection The human MM A375 cell line was purchased from the Biovector Science Lab, Inc. (Beijing, China).

Techniques: Plasmid Preparation

Journal: Experimental and Therapeutic Medicine

Article Title: The biological effect and mechanism of the Wnt/β-catenin signaling pathway on malignant melanoma A375 cells

doi: 10.3892/etm.2018.6413

Figure Lengend Snippet: Influence of lentiviral vector-mediated siRNA on A375 cell invasion.

Article Snippet: Cell culture and transfection The human MM A375 cell line was purchased from the Biovector Science Lab, Inc. (Beijing, China).

Techniques: Plasmid Preparation

Journal: Experimental and Therapeutic Medicine

Article Title: The biological effect and mechanism of the Wnt/β-catenin signaling pathway on malignant melanoma A375 cells

doi: 10.3892/etm.2018.6413

Figure Lengend Snippet: Influence of lentiviral vector-mediated siRNA on A375 cell (A) invasion and (B) migration in vitro. siRNA, small interfering RNAm (magnification, ×200).

Article Snippet: Cell culture and transfection The human MM A375 cell line was purchased from the Biovector Science Lab, Inc. (Beijing, China).

Techniques: Plasmid Preparation, Migration, In Vitro

Journal: Experimental and Therapeutic Medicine

Article Title: The biological effect and mechanism of the Wnt/β-catenin signaling pathway on malignant melanoma A375 cells

doi: 10.3892/etm.2018.6413

Figure Lengend Snippet: Influence of lentiviral vector-mediated siRNA on A375 cell migration.

Article Snippet: Cell culture and transfection The human MM A375 cell line was purchased from the Biovector Science Lab, Inc. (Beijing, China).

Techniques: Plasmid Preparation, Migration

Journal: Experimental and Therapeutic Medicine

Article Title: The biological effect and mechanism of the Wnt/β-catenin signaling pathway on malignant melanoma A375 cells

doi: 10.3892/etm.2018.6413

Figure Lengend Snippet: Influence of lentiviral vector-mediated siRNA on A375 cell apoptosis.

Article Snippet: Cell culture and transfection The human MM A375 cell line was purchased from the Biovector Science Lab, Inc. (Beijing, China).

Techniques: Plasmid Preparation

Journal: Genes

Article Title: Integrative Transcriptomic Analysis Identify Potential m6A Pathway-Related Drugs That Inhibit Cancer Cell Proliferation

doi: 10.3390/genes13112011

Figure Lengend Snippet: Effect of R428 on human melanoma cell line cell line A375. ( A , B ) The proliferation of the human breast cancer cell line A375 was significantly inhibited by R428 treatment. The results from the cell count assay ( A ) and the MTT assay ( B ) are shown. Data represented the mean ± SEM from five independent experiments, t -test, * p < 0.05, ** p < 0.01. Some SEM values in the drug-treated experimental group that are too small to be shown are specified here: ( A ) 24 h. 0.009; 48 h, 0.012; 72 h, 0.006; 96 h, 0.000; (B) 24 h. 0.004; 48 h, 0.002; 72 h, 0.006; 96 h, 0.002. ( C , D ) The protein expression of METTL3 in the A375 cell line was significantly reduced by drug treatment, t -test, ** p < 0.01. The summary of Western blot grey values as the mean ± SEM from three independent experiments ( C ) and the representative Western blot result ( D ) are shown. ( E ) The RNA m6A modification level in the A375 cell line was significantly decreased after R428 treatment. Data represented the mean ± SEM from five independent experiments, t -test, * p < 0.05. ( F , G ) Representative images of treated and control cells observed under an inverted 10× microscopy.

Article Snippet: The breast cancer cell line MCF7 and the human melanoma cell line A375 were provided by Biomol Inc. (Hamburg, Germany).

Techniques: Cell Counting, MTT Assay, Expressing, Western Blot, Modification, Control, Microscopy