two-compartment boyden chamber matrigel invasion assay costar transwell  (Corning Life Sciences)

Journal: Cancer Medicine

Article Title: S -adenosylmethionine blocks osteosarcoma cells proliferation and invasion in vitro and tumor metastasis in vivo : therapeutic and diagnostic clinical applications

doi: 10.1002/cam4.386

Figure Lengend Snippet: Effect of S -adenosylmethionine (SAM) on osteosarcoma (OS) cells proliferation and invasion in vitro . Human OS cancer cells LM-7 and MG-63 were plated in 10 mL plates and treated with 150 μ mol/L of S -adenosylhomocystine as control (SAH) or two doses (75 and 150 μ mol/L) of SAM. Cell growth rate was determined in each group by trypsinization and counting the number of cells by Coulter counter as described in Materials and Methods section (A). LM-7 and MG-63 cells invasive capacity was evaluated by using a Boyden chamber Matrigel invasion assay. After 18 h of SAM (75 and 150 μ mol/L) treatment, the invaded cells were fixed, stained, and 10 random fields were counted. Number of cells invading is shown as bar diagram ± SEM (B) as described in Materials and Methods section. Results are presented as the mean ± SEM of two different experiments in duplicate from control and experimental cells. Significant differences from the control (SAH) is represented by an asterisk ( P < 0.05).

Article Snippet: The invasive capacity of LM-7 and MG-63 cells was examined using two-compartment Boyden chamber Matrigel invasion assay (Costar Transwell, Corning Corporation, Sigma-Aldrich, Oakville, ON, Canada) following treatment with SAH or SAM for 6 days as described previously .

Techniques: In Vitro, Control, Invasion Assay, Staining

Journal: Cancer Medicine

Article Title: LRP5 knockdown: effect on prostate cancer invasion growth and skeletal metastasis in vitro and in vivo

doi: 10.1002/cam4.111

Figure Lengend Snippet: Effect of DN-LRP5 on PC-3 cells invasion, colony formation, and migration in vitro . Control (PC-3-pcDNA3.1) and experimental (PC-3-DN-LRP5) cells invasive capacity was evaluated by using a Boyden chamber Matrigel invasion assay. After 18 h of incubation, the invaded cells were fixed, stained, and 10 random fields were counted. Number of cells invading is shown as bar diagram ± SEM (A) as described in “Material and Methods”. Using soft agar assay, the ability of control and experimental cells to form colonies was examined and the number of colonies formed in each group was counted (B) as described in “Material and Methods”. PC-3 cells migration was determined by wound healing assay by seeding PC-3-pcDNA3.1 and PC-3-DN-LRPP5 cells in six-well plates and allowing them to grow as a monolayer and making a wound as described in “Material and Methods”. Control and experimental migrating cells were photographed at different time points (C). Percent wound healing was recorded at different time points, and percentage of wound healing with respect to T 0 was calculated using the equation described in “Material and Methods”. Results are presented as the mean ± SEM of two different experiments in duplicate from control and experimental cells. Significant differences from the control is represented by an asterisk ( P < 0.05).

Article Snippet: The invasive capacity of 5 × 10 4 PC-3-pcDNA3.1 and PC-3-DN-LRP5 cells were examined using two-compartment Boyden chamber Matrigel invasion assay (Costar Transwell, Corning Corporation, Corning, NY) as described previously .

Techniques: Migration, In Vitro, Invasion Assay, Incubation, Staining, Soft Agar Assay, Wound Healing Assay

Journal:

Article Title: Pharmacological Inhibition of DNA Methylation Induces Proinvasive and Prometastatic Genes In Vitro and In Vivo 1

doi:

Figure Lengend Snippet: Effect of 5-aza-CdR treatment on the proliferation, invasion, and migration of noninvasive cell lines. Noninvasive breast cancer cells, MCF-7 and ZR-75-1, were seeded at the same density and subjected to two different doses of 5-aza-CdR or with vehicle control alone (CTL) in a six-well plate, and growth curve analysis was carried out to determine the effect of 5-aza-CdR on the proliferation rate. At each time point, cells were trypsinized and counted as described in the Materials and Methods section (A and B). All data presented are expressed as mean ± SEM of quadruplicate wells for each time point. Similar results were obtained in two independent experiments. Cell cycle distribution analysis was carried out by FACS for control and 5-aza-CdR-treated MCF-7 and ZR-75-1 cells (5 µM for 7 days) after staining the cells with propidium iodide as described in the Materials and Methods section. Data were analyzed by WinMDI version 2.8 and plotted (C). Data are presented as mean of % events from three independent experiments by 100% stacked column graph for control and experimental groups. Expression of uPA, a protease involved in invasion, was analyzed by RT-PCR in MCF-7 and ZR-75-1 cells treated with different doses of 5-aza-CdR (D). Data were presented for control and experimental groups as mean ± SEM values from two independent experiments. The invasive capacity of MCF-7 and ZR-75-1 cells treated with different doses of 5-aza-CdR was determined using a Boyden chamber Matrigel invasion assay. The bar diagram represents the mean ± SEM as described in the Materials and Methods section. Wound healing migration assay was carried out on MCF-7 cells in the absence and presence of 5-aza-CdR (1 and 5µ M) in the culture medium with 2% fetal bovine serum as depicted in panels E and F. Cells were seeded at the same density and grown as monolayer and wounded as discussed in the Materials and Methods section. % Wound healing was recorded at different time points, and percentage of wound healing with respect to T0 was calculated using the described equation. Similar results were obtained from two different experiments. All data were presented as mean ± SEM of duplicate values from control and experimental groups from two independent experiments. Significant difference from the control is represented by an asterisk (*P < .05).

Article Snippet: Cells' invasive capacity was determined using a two-compartment Boyden chamber Matrigel invasion assay (Transwell; Costar Corning Corporation, Burlington, MA) as described previously [ 21 ].

Techniques: Migration, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Invasion Assay