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: 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: Frontiers in Oncology

Article Title: TRAF3IP3 promotes glioma progression through the ERK signaling pathway

doi: 10.3389/fonc.2022.776834

Figure Lengend Snippet: TRAF3IP3 promotes tumor progression via ERK signaling. (A) TRAF3IP3 knockdown in U251 cells decreased the phosphorylation of ERK, whereas TRAF3IP3 overexpression in U118 cells increased the phosphorylation of ERK. (B) TRAF3IP3 overexpression of U118 cells increased ERK activation. (C,D) CCK-8 assay and EdU assay showed that TRAF3IP3 activated the ERK signaling pathway to promote tumor proliferation, which could be rescued by U0126. (E) Wound-healing assay showed that TRAF3IP3 activated the ERK signaling pathway to promote tumor migration, which could be rescued by U0126. (F) Transwell assays showed that TRAF3IP3 activated the ERK signaling pathway to promote tumor invasion, which could be rescued by U0126. (G) The effect of TRAF3IP3 overexpression in activating ERK signaling pathway could be rescued by U0126. *p < 0.05, **p<0.01, ***p<0.001, ### p<0.001.

Article Snippet: Transwell invasion assays were performed using Transwell chambers with Matrigel (354480, CORNING).

Techniques: Knockdown, Phospho-proteomics, Over Expression, Activation Assay, CCK-8 Assay, EdU Assay, Wound Healing Assay, Migration

Journal: Journal of Cancer

Article Title: A Positive Feedback Loop of E2F4-Mediated Activation of MNX1 Regulates Tumour Progression in Colorectal Cancer

doi: 10.7150/jca.86718

Figure Lengend Snippet: MNX1 knockdown affects the migration and invasion of colorectal cancer cells. (A, B) MNX1 mRNA and protein levels are downregulated in cell lines; (C) MTT analysis of the HCT116 and SW620 cells for the different groups; (D, E) Wound healing assays showed that MNX1 downregulation significantly reduced cell migration; (F, G) The results of the transwell assay revealed that MNX1 downregulation in HCT116 and SW620 cells decreased cell invasion and migration. Mean ± SD (n = 3 independent experiments). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Cell migration and invasion assays were conducted in 6-well Transwell chambers (8 μm filter pore, Corning) pre-coated or not precoated with Matrigel basement membrane gel.

Techniques: Knockdown, Migration, Transwell Assay

Journal: Journal of Cancer

Article Title: A Positive Feedback Loop of E2F4-Mediated Activation of MNX1 Regulates Tumour Progression in Colorectal Cancer

doi: 10.7150/jca.86718

Figure Lengend Snippet: MNX1 overexpression affects the migration and invasion of colorectal cancer cells. (A, B) Upregulated MNX1 mRNA and protein levels in cell lines; (C) The MTT assay results showed the proliferation of the HCT116 and SW620 cells for different groups; (D, E) Wound healing assays revealed that MNX1 upregulation substantially promoted cell migration; (F, G) The results of the transwell assay showed that MNX1 upregulation in HCT116 and SW620 cells increased cell invasion and migration. Mean ± SD (n = 3 independent experiments). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Cell migration and invasion assays were conducted in 6-well Transwell chambers (8 μm filter pore, Corning) pre-coated or not precoated with Matrigel basement membrane gel.

Techniques: Over Expression, Migration, MTT Assay, Transwell Assay