Journal: Cureus

Article Title: The Role of Matrix Metalloproteinase-2 (MMP2) in Colorectal Cancer Progression: Correlation With Clinicopathological Features and Impact on Cellular Processes

doi: 10.7759/cureus.61941

Figure Lengend Snippet: Correlation between MMP2 expression and clinicopathological features of CRC patients T: tumor depth; N: lymph node involvement; M: metastasis; MMP2: matrix metalloproteinase-2; CRC: colorectal cancer * Based on Chi-square test, or (if not applicable) Fisher’s exact test

Article Snippet: Subsequently, the dishes were further incubated with 5 μM of the MMP2 inhibitor ARP100 (MMP-2 inhibitor from Santa Cruz Biotechnology, Dallas, USA), dissolved in dimethyl sulfoxide (DMSO) at the specified concentration as needed for the experiments.

Techniques: Expressing

Journal: Cureus

Article Title: The Role of Matrix Metalloproteinase-2 (MMP2) in Colorectal Cancer Progression: Correlation With Clinicopathological Features and Impact on Cellular Processes

doi: 10.7759/cureus.61941

Figure Lengend Snippet: Differences in MMP2 expression between cancerous and cancerous-adjacent normal tissues * Based on Chi-square test, or (if not applicable) Fisher’s exact test; MMP2: matrix metalloproteinase-2

Article Snippet: Subsequently, the dishes were further incubated with 5 μM of the MMP2 inhibitor ARP100 (MMP-2 inhibitor from Santa Cruz Biotechnology, Dallas, USA), dissolved in dimethyl sulfoxide (DMSO) at the specified concentration as needed for the experiments.

Techniques: Expressing

Journal: Cureus

Article Title: The Role of Matrix Metalloproteinase-2 (MMP2) in Colorectal Cancer Progression: Correlation With Clinicopathological Features and Impact on Cellular Processes

doi: 10.7759/cureus.61941

Figure Lengend Snippet: The representative image showed elevated MMP2 expression levels in (A) cancerous tissues compared to those in (B) normal tissues (20x). MMP2: matrix metalloproteinase-2

Article Snippet: Subsequently, the dishes were further incubated with 5 μM of the MMP2 inhibitor ARP100 (MMP-2 inhibitor from Santa Cruz Biotechnology, Dallas, USA), dissolved in dimethyl sulfoxide (DMSO) at the specified concentration as needed for the experiments.

Techniques: Expressing

Journal: Cureus

Article Title: The Role of Matrix Metalloproteinase-2 (MMP2) in Colorectal Cancer Progression: Correlation With Clinicopathological Features and Impact on Cellular Processes

doi: 10.7759/cureus.61941

Figure Lengend Snippet: MMP2 was inhibited in SW480 cells by treating with 5 μM of the MMP2 inhibitor ARP100 for 24h, while control cells were treated with DMSO. (A-C) Transwell migration and invasion assays (magnification: 100×) demonstrated that MMP2 inhibition significantly reduced the migratory and invasive capabilities of SW480 cells. (D and E) Wound healing assays (scale: 100 µm) showed that MMP2 inhibition markedly decreased gap closure in SW480 cells. Data are presented as mean ± standard error (**P≤0.002; ***P<0.0005). (F) Western blot analysis confirmed MMP2 inhibition. Results are representative of three independent experiments. MMP2: matrix metalloproteinase-2; CRC: colorectal cancer; DMSO: dimethyl sulfoxide

Article Snippet: Subsequently, the dishes were further incubated with 5 μM of the MMP2 inhibitor ARP100 (MMP-2 inhibitor from Santa Cruz Biotechnology, Dallas, USA), dissolved in dimethyl sulfoxide (DMSO) at the specified concentration as needed for the experiments.

Techniques: Control, Migration, Inhibition, Western Blot

Journal: Cureus

Article Title: The Role of Matrix Metalloproteinase-2 (MMP2) in Colorectal Cancer Progression: Correlation With Clinicopathological Features and Impact on Cellular Processes

doi: 10.7759/cureus.61941

Figure Lengend Snippet: MMP2 was inhibited in SW480 cells by treating with 5 μM of the MMP2 inhibitor ARP100, while control cells were treated with DMSO. Cell proliferation was assessed using the MTT assay, with absorbance readings taken at 570 nm. (A) APR100 inhibitor significantly inhibited SW480 cell proliferation. (B and C) Transfection efficiency was validated through RT-PCR and western blot analysis. Data are presented as mean ± standard error (*P<0.02 and **P≤0.009). Results from three representative experiments are depicted. MMP2: matrix metalloproteinase-2; DMSO: dimethyl sulfoxide; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide; RT-PCR: reverse transcription-polymerase chain reaction

Article Snippet: Subsequently, the dishes were further incubated with 5 μM of the MMP2 inhibitor ARP100 (MMP-2 inhibitor from Santa Cruz Biotechnology, Dallas, USA), dissolved in dimethyl sulfoxide (DMSO) at the specified concentration as needed for the experiments.

Techniques: Control, MTT Assay, Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot, Reverse Transcription, Polymerase Chain Reaction

Journal: Cureus

Article Title: The Role of Matrix Metalloproteinase-2 (MMP2) in Colorectal Cancer Progression: Correlation With Clinicopathological Features and Impact on Cellular Processes

doi: 10.7759/cureus.61941

Figure Lengend Snippet: Caspase 3 and 9 activities in SW480 treated cells were quantified using spectrophotometry. The activity of caspases in control cells (treated with DMSO) was set as 1, and the activity values of the experimental group were standardized relative to this. (A) SW480 cells treated with inhibitor exhibited a significant elevation in both caspase 3 and caspase 9 activities. (B) Western blot analysis was performed to detect the expression levels of apoptosis-related proteins. Bar graphs depict the standard errors of the mean from three experiments (**P≤0.005 or P=0.001). MMP2: matrix metalloproteinase-2; DMSO: dimethyl sulfoxide

Article Snippet: Subsequently, the dishes were further incubated with 5 μM of the MMP2 inhibitor ARP100 (MMP-2 inhibitor from Santa Cruz Biotechnology, Dallas, USA), dissolved in dimethyl sulfoxide (DMSO) at the specified concentration as needed for the experiments.

Techniques: Spectrophotometry, Activity Assay, Control, Western Blot, Expressing

Journal: Frontiers in Immunology

Article Title: Arsenic exposure and lung fibrotic changes-evidence from a longitudinal cohort study and experimental models

doi: 10.3389/fimmu.2023.1225348

Figure Lengend Snippet: MMP-2 is critical for arsenic-induced EMT changes. (A) NHBE cells were pretreated with ARP100 2 hours before NaAsO 2 treatment. After another 24 hours of combined treatment, the cells were applied for western blot analysis and wound healing assay. ARP-100 reverse NaAsO 2 -induced mesenchymal marker expressions (A) and cell migration (B) . Each result was performed in three independent experiments. The data were expressed as Mean+/-SEM. a: p <0.05 compared to untreated control; b: p <0.05 compared to NaAsO 2 group.

Article Snippet: ARP-100 was purchased from Santa Cruz (Santa Cruz, CA).

Techniques: Western Blot, Wound Healing Assay, Marker, Migration

Journal: bioRxiv

Article Title: Formate Promotes Invasion and Metastasis by Activating Fatty Acid Synthesis and Matrix Metalloproteinases

doi: 10.1101/2023.01.23.525172

Figure Lengend Snippet: (A) Invasion of LN-229 cells treated for 24 hours with 500 μM Na-formate was assed on differently coated Boyden chambers (Collagen, ECM, Fibronectine, Laminin, PLL, and ECM-Collagen). Each dot represents an independent experiment; mean ± SD; Unpaired t -test with Welch’s correction. (B) RNAseq results displaying the count of different MMPs (MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP11, MMP13, MMP14, MMP15, MMP16, MMP17, MMP19, MMP23, MMP24, MMP25, and MMP28) expression in LN-229 cells which were treated for 8 hours or 24 hours with 500 μM Na-formate. Each dot represents an independent experiment; mean ± SD; Unpaired t -test with Welch’s correction. mRNA expression of MMP2, MMP9, MMP11, and MMP16 in LN-229 and U87 cells which were treated for 8 h or 24 h with 500 μM Na-formate relative to untreated cells as measured using real-time RT-qPCR. Each dot represents an independent experiment; mean ± SD. (D) Invasion of LN-229 cells treated for 24 h with 500 μM formate and 5 μM MMP inhibitors (MMP2 inhibitor and MMP9 inhibitor) was assed using ECM-Collagen coated Boyden chambers. Each dot represents an independent experiment; mean ± SD; Unpaired t -test with Welch’s correction. (E) Invasion of U87 cells treated for 24 h with 500 μM formate and 5 μM MMP inhibitors (MMP2 inhibitor and MMP9 inhibitor) was assed using ECM-Collagen coated Boyden chambers. Each dot represents an independent experiment; mean ± SD; Unpaired t -test with Welch’s correction. (F) Displacement of LN-229 cells injected in ex vivo brain slices. The cells were cultured during the entire experiment with 500 μM formate, 5 μM ARP100, or the combination of both. Results are displayed as mean ± SEM; Unpaired t -test with Welch’s correction. Each dot represent the average displacement of 15 cells.

Article Snippet: The MMP inhibitors: ARP100 (MMP2 inhibitor) ( Santa Cruz: SC-203522 ) and MMP-9 Inhibitor I (MMP9 inhibitor) ( Santa Cruz: SC-311437 ) were purchased from Santa Cruz.

Techniques: Expressing, Quantitative RT-PCR, Injection, Ex Vivo, Cell Culture

Journal: bioRxiv

Article Title: Formate Promotes Invasion and Metastasis by Activating Fatty Acid Synthesis and Matrix Metalloproteinases

doi: 10.1101/2023.01.23.525172

Figure Lengend Snippet: (A) Migration of different cells: BG5, BG7, GG16, LN-229, U87, and MDA-MB-468 treated for 24 hours with 500 μM formate was assed using non-coated Boyden chambers. Each dot represents an independent experiment; mean ± SD; Unpaired t -test with Welch’s correction. (B) Migratory potential of LN-229 cells in response to different formate concentrations (0 μM, 50 μM, 100 μM, 200 μM, 500 μM, and 1mM) as time-dependent quantification of relative wound density in IncuCyte live cell analysis. Graph shows mean ± SEM ( n = 3). (C) mRNA expression from MMP2, MMP9, MMP11, and MMP16 in BT20, 4T1, MDA-MB-468, and MDA-MB-231 cells which were treated for 24 hours with 500 μM formate relative to untreated cells as measured using real-time RT-qPCR. Each dot represents an independent experiment; mean ± SD. (D) Expression of MMP2 or MMP9 in the cell culture medium. The total stain served as a loading control. The signal intensity of MMP has been quantified with respect to the signal intensity of the total stain. Each dot represents an independent experiment, the bars represent the mean, and the error bars visualize the standard deviation. The data was evaluated using an unpaired t -test with Welch’s correction. (E) MMP2 and MMP9 activity of LN-229 sh Scramble or MTHFD1L KD cells treated for 72 hours with 500 μM formate was assed using gelatin zymography assay. Each dot represents an independent experiment; mean ± SD; Unpaired t -test with Welch’s correction. (F) Proliferation of LN-229 cells treated for 72 h with different concentrations (0 μM, 1 μM, 2.5 μM, 5 μM, and 10 μM) of MMP2 or MMP9 inhibitor was determined as a time-dependent quantification of cell density (confluence) in IncuCyte. Results are displayed as a fold change to time point zero ( n = 3).

Article Snippet: The MMP inhibitors: ARP100 (MMP2 inhibitor) ( Santa Cruz: SC-203522 ) and MMP-9 Inhibitor I (MMP9 inhibitor) ( Santa Cruz: SC-311437 ) were purchased from Santa Cruz.

Techniques: Migration, Expressing, Quantitative RT-PCR, Cell Culture, Staining, Standard Deviation, Activity Assay, Zymography Assay