Journal: Heliyon

Article Title: Germinal center B-cell subgroups in the tumor microenvironment cannot be overlooked: Their involvement in prognosis, immunotherapy response, and treatment resistance in head and neck squamous carcinoma

doi: 10.1016/j.heliyon.2024.e37726

Figure Lengend Snippet: | PLXNB2 significantly affects the proliferation and migration of head and neck cancer cell lines. (A, B) CCK-8 experiment. After PLXNB2 knockdown, the proliferation ability of HN-5 and UMSCC-47 cell lines decreased significantly. (C, D) Plate cloning experiment. After PLXNB2 knockdown, the colony-forming ability of HN-5 and UMSCC-47 cell lines was significantly decreased. (E, F) wound healing test. After PLXNB2 knockdown, the migration ability of HN-5 and UMSCC-47 cell lines decreased significantly. (G, H) Transwell experiment. After PLXNB2 knockdown, the migration and invasion ability of HN-5 and UMSCC-47 cell lines were significantly reduced. (∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001).

Article Snippet: In the Transwell experimental setup, the Transwell chamber (Corning, USA, 341201) was positioned within a 24-well plate.

Techniques: Migration, CCK-8 Assay, Knockdown, Cloning

Journal: Cancer Science

Article Title: Mesenchymal-transitioned cancer cells instigate the invasion of epithelial cancer cells through secretion of WNT3 and WNT5B

doi: 10.1111/cas.12336

Figure Lengend Snippet: Cell–cell contact independent induction of secondary epithelial-to-mesenchymal transition (EMT) phenotype and invasiveness in neighboring epithelial cancer cells by mesenchymal-transitioned cancer cells. (a, b) A549 cells were co-cultured directly (a) or separately in transwell cell culture chamber (b) at the indicated cell numbers for 24 h and EMT-related protein expression were determined by western blotting. No cells (−), E-cells (E) or M-cells (M) were seeded in upper compartment of transwell chamber. In the separated co-culture, the total protein from E-cells in lower compartment was examined. (c, d) Epithelial A549 cells were treated with conditioned mediums (CM) from E-/M-A549 cells (c) or E-/M-Panc cells (d) for 48 h and subjected to Matrigel invasion assay or western blotting. Total invaded cells were counted after H&E staining. Data represented as the mean ± SD of triplicate experiment. ** P < 0.01 versus E-CM group by two-tailed Student's t test.

Article Snippet: For the direct co-culture experiment, E-cells and M-cells were re-seeded into 60 mm culture dish according to the indicated cell number and co-cultured for 24 h. For the separated co-culture experiment, 1 × 10 5 E-cells and 3 × 10 5 cells of M-cells were seeded into lower or upper compartments of the transwell chamber with 1 μm pore diameter (BD Falcon, Bedford, MA, USA) for 24 h.

Techniques: Cell Culture, Expressing, Western Blot, Co-Culture Assay, Invasion Assay, Staining, Two Tailed Test

Journal: Experimental and Therapeutic Medicine

Article Title: miR-124 inhibits proliferation, migration and invasion of malignant melanoma cells via targeting versican

doi: 10.3892/etm.2017.4998

Figure Lengend Snippet: A375 cells were transfected with miR-NC or miR-124 mimics. (A) Reverse-transcription quantitative polymerase chain reaction analysis was used to examine the expression of miR-124. (B) MTT assay, (C) wound healing assay (magnification, ×40) and (D) Transwell assay (magnification, ×200) were used to examine cell proliferation, migration and invasion, respectively. **P<0.01 vs. miR-NC. miR, microRNA; miR-NC, scrambled negative control miR mimics.

Article Snippet: A Transwell assay was performed to detect the cell invasion capacity using Transwell chambers (BD Biosciences, Franklin Lakes, NJ, USA) coated with Matrigel.

Techniques: Transfection, Real-time Polymerase Chain Reaction, Expressing, MTT Assay, Wound Healing Assay, Transwell Assay, Migration, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: miR-124 inhibits proliferation, migration and invasion of malignant melanoma cells via targeting versican

doi: 10.3892/etm.2017.4998

Figure Lengend Snippet: miR-124-overexpressing A375 cells were transfected with pcDNA3.1-versican open reading frame plasmid or blank pcDNA.31 vector as an NC. (A) Reverse-transcription quantitative polymerase chain reaction analysis and (B) western blot were used to assess the mRNA and protein expression of versican, respectively. (C) MTT assay, (D) wound healing assay (magnification, ×40) and (E) Transwell assay (magnification, ×200) were used to examine cell proliferation, migration and invasion, respectively. **P<0.01 vs. miR-124+NC. NC, negative control; miR, microRNA.

Article Snippet: A Transwell assay was performed to detect the cell invasion capacity using Transwell chambers (BD Biosciences, Franklin Lakes, NJ, USA) coated with Matrigel.

Techniques: Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Western Blot, Expressing, MTT Assay, Wound Healing Assay, Transwell Assay, Migration, Negative Control

Journal: Yonsei Medical Journal

Article Title: Heat Shock Factor 1 Predicts Poor Prognosis of Gastric Cancer

doi: 10.3349/ymj.2018.59.9.1041

Figure Lengend Snippet: Downregulation of HSF1 expression reduces proliferation, migration, and invasion of gastric cancer cells. AGS and MKN28 cells were transfected with a scrambled siRNA (scRNA) or two small-interfering RNAs (siRNAs) specific for HSF1 (siRNA #1 and #2). (A) HSF1 protein expression was detected by Western blot analysis. β-actin was used as a loading control. (B) WST assay was performed to detect cell viability. (C and D) Transwell assays to evaluate (C) migration and (D) invasion of cells (×200). The histogram is represented as mean±SEM (n=3). p values were calculated using ANOVA and statistically significant differences are indicated as * ( * p <0.001). HSF1, heat shock factor 1.

Article Snippet: After 24-hour transfection, 1×10 4 cells from each well were isolated and added to the upper transwell chamber (Corning Costar, Tewksbury, MA, USA) that carried a filter coated with 0.5 mg/mL of collagen type I (BD Biosciences, Seoul, Korea) for the migration assay or a filter coated with Matrigel (1:15) (BD Biosciences) for the invasion assay.

Techniques: Expressing, Migration, Transfection, Western Blot, Control, WST Assay

Journal: Yonsei Medical Journal

Article Title: Heat Shock Factor 1 Predicts Poor Prognosis of Gastric Cancer

doi: 10.3349/ymj.2018.59.9.1041

Figure Lengend Snippet: Overexpression of HSF1 promotes proliferation, migration, and invasion of gastric cancer cells. Overexpression of HSF1 in AGS and MKN28 cells was achieved by transfection with an empty vector (pcDNA_EV) or HSF1 overexpression vector (pcDNA_HSF1). (A) Expressions of HSF1 mRNA and protein were detected by Western blot analysis. β-actin was used as a loading control. (B) WST assay was performed to detect cell viability. (C and D) Transwell migration assays to evaluate (C) migration and (D) invasion activity of cells (×200). Data are represented as mean±SEM (n=3). p values were calculated using Student's t-test ( * p <0.001, † p =0.001). HSF1, heat shock factor 1.

Article Snippet: After 24-hour transfection, 1×10 4 cells from each well were isolated and added to the upper transwell chamber (Corning Costar, Tewksbury, MA, USA) that carried a filter coated with 0.5 mg/mL of collagen type I (BD Biosciences, Seoul, Korea) for the migration assay or a filter coated with Matrigel (1:15) (BD Biosciences) for the invasion assay.

Techniques: Over Expression, Migration, Transfection, Plasmid Preparation, Western Blot, Control, WST Assay, Activity Assay

Journal: OncoTargets and therapy

Article Title: miR-495 inhibits proliferation, migration, and invasion and induces apoptosis via inhibiting PBX3 in melanoma cells

doi: 10.2147/OTT.S152362

Figure Lengend Snippet: miR-495 inhibits melanoma cell proliferation, migration, invasion, and colony formation in vitro. ( A ) A375 and MeWo cells were transfected with miR-495 mimics, negative control (miR-NC), negative control inhibitor (NC inhibitor), or miR-495 inhibitor for 24 hours, then the expression of miR-495 was quantified by qRT-PCR. ( B ) miR-495 mimics, miR-NC, NC inhibitor or miR-495 inhibitor-transfected A375, and MeWo cells were subjected to MTT assay. ( C ) miR-495 mimics, miR-NC, NC inhibitor or miR-495 inhibitor-transfected A375, and MeWo cells were subjected to wound healing assay. ( D ) miR-495 mimics, miR-NC, NC inhibitor or miR-495 inhibitor-transfected A375, and MeWo cells were subjected to transwell assay. ( E ) miR-495 mimics, miR-NC, NC inhibitor or miR-495 inhibitor-transfected A375, and MeWo cells were subjected to colony formation assay. Data are presented as mean ± SD (n=3). All experiments were performed three times, and representative images are presented. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: About 1×10 5 cells were plated on the upper Transwell chambers (Costar; Corning Incorporated, Corning, NY, USA) with Matrigel (BD Bioscience, San Jose, CA, USA).

Techniques: Migration, In Vitro, Transfection, Negative Control, Expressing, Quantitative RT-PCR, MTT Assay, Wound Healing Assay, Transwell Assay, Colony Assay

Journal: OncoTargets and therapy

Article Title: miR-495 inhibits proliferation, migration, and invasion and induces apoptosis via inhibiting PBX3 in melanoma cells

doi: 10.2147/OTT.S152362

Figure Lengend Snippet: Silencing of pre-B-cell leukemia transcription factor 3 (PBX3)-inhibited cell proliferation, migration, and invasion in melanoma cells. ( A ) A375 and MeWo cells were transfected with negative control siRNA (si-NC) or siRNA against PBX3 (si-PBX3) for 24 hours, the mRNA expression levels of PBX3 in both the cells were examined by RT-qPCR and ( B ) the protein levels of PBX3 in both the cells were examined by Western blot. ( C ) A375 and MeWo cells were transfected with si-NC or si-PBX3 for indicated time, and cell proliferation was analyzed by MTT assay. ( D ) A375 and MeWo cells were transfected with si-NC or by transwell assay. ( E ) Cell migration was analyzed by si-PBX3 for 24 hours, and cell invasion was analyzed by wound healing assay. Data are presented as mean ± SD (n=3). All experiments were performed three times, and representative images are presented. * P <0.05, ** P <0.01, *** P <0.001. Abbreviation: NC, negative control.

Article Snippet: About 1×10 5 cells were plated on the upper Transwell chambers (Costar; Corning Incorporated, Corning, NY, USA) with Matrigel (BD Bioscience, San Jose, CA, USA).

Techniques: Migration, Transfection, Negative Control, Expressing, Quantitative RT-PCR, Western Blot, MTT Assay, Transwell Assay, Wound Healing Assay

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Overexpressed MPS-1 contributes to endometrioma development through the NF-κB signaling pathway

doi: 10.1186/s12958-021-00796-z

Figure Lengend Snippet: MPS-1 regulates proliferation, migration, invasion, apoptosis and cell cycle of EcESCs. A Transfection efficiency of MPS-1 siRNA was detected by qRT-PCR. B A representative western blotting image of MPS-1 after EcESCs were transfected with MPS-1 siRNA for 72 h (n = 2). C Based on the results of western blotting, knockdown efficiency of MPS-1 siRNA was analyzed by Image J software. D The effects of MPS-1 siRNA on proliferation were evaluated by CCK-8 assay. E , G The representative images of EcESCs apoptosis and cell cycle were depicted by flow cytometry after MPS-1 siRNA transfection for 72 h. F , H The apoptosis analysis and cell cycle analysis were based on the results of flow cytometry. I , K Transwell assays were used to determine the effects of MPS-1 siRNA on migration and invasion. The scale bar is 50 μm. J , L Cell count analysis of Transwell migration and invasion. NC, si-MPS-1 EcESCs transfected with negative control siRNA or MPS-1 siRNA (n = 4). Data are presented as mean ± SEM (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)

Article Snippet: After 48 h of transfection, 5 × 10 4 EcESCs in 200 μL DMEM/F12 were seeded onto the upper transwell chamber (BD Falcon, USA).

Techniques: Migration, Transfection, Quantitative RT-PCR, Western Blot, Knockdown, Software, CCK-8 Assay, Flow Cytometry, Cell Cycle Assay, Cell Counting, Negative Control