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

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Warburg effect-promoted exosomal circ_0072083 releasing up-regulates NANGO expression through multiple pathways and enhances temozolomide resistance in glioma

doi: 10.1186/s13046-021-01942-6

Figure Lengend Snippet: circ_0072083 silence decreases TMZ resistance in the resistant glioma cells. a circ_0072083 expression was detected via qRT-PCR in resistant cells transfected with sh-circ_0072083 or sh-NC. b IC50 of TMZ was analyzed via CCK-8 in cells with transfection of sh-circ_0072083 or sh-NC. Cell proliferation c , colony ability d , apoptosis e , migration f and g and invasion h were measured by CCK-8, colony formation, flow cytometry, wound healing and transwell analyses respectively in resistant cells transfected with sh-circ_0072083 or sh-NC after treatment of 50 μg/mL of TMZ. i and j Tumor volume and weight were detected in each group ( n = 5). Xenograft model was established using U251/TR cells transfected with sh-circ_0072083 or sh-NC, and then treated via PBS or TMZ. k circ_0072083 abundance was measured via qRT-PCR in tumor tissues of each group. * P < 0.05

Article Snippet: For transwell migration analysis, 24-well transwell chambers (Corning Inc., Corning, NY, USA) with a fibronectin-coated polycarbonate membrane were used.

Techniques: Expressing, Quantitative RT-PCR, Transfection, CCK-8 Assay, Migration, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Warburg effect-promoted exosomal circ_0072083 releasing up-regulates NANGO expression through multiple pathways and enhances temozolomide resistance in glioma

doi: 10.1186/s13046-021-01942-6

Figure Lengend Snippet: circ_0072083 targets miR-1252-5p to regulate ALKBH5/NANOG and TMZ resistance in the resistant glioma cells. a The potential miRNA that interacted with circ_0072083, ALKBH5 and NANOG was analyzed via Circinteractome and starBase. b The binding site of miR-1252-5p and circ_0072083, ALKBH5 and NANOG was exhibited. c miR-1252-5p expression were detected via qRT-PCR in TMZ-resistant ( n = 36) or sensitive tissues ( n = 33). d miR-1252-5p abundance was measured by qRT-PCR in TME-resistant and sensitive cells. e - g Luciferase activity was detected in 293 T cells with transfection of miR-1252-5p mimic or miR-NC and the constructed luciferase reporter vectors. h miR-1252-5p, circ_0072083, ALKBH5 and NANOG levels were detected after RIP assay using anti-Ago2 or anti-IgG. i miR-1252-5p expression was measured via qRT-PCR in cells transfected with sh-circ or sh-NC. (J and K) ALKBH5 and NANOG levels were detected via qRT-PCR and western blot in cells transfected with miR-1252-5p mimic or miR-NC. l miR-1252-5p expression was measured via qRT-PCR in cells transfected with anti-miR-1252-5p or anti-NC. m and n ALKBH5 and NANOG levels were examined via qRT-PCR and western blot in cells transfected with sh-NC + anti-NC, sh-circ_0072083 + anti-NC or sh-circ_0072083 + anti-miR-1252-5p. IC50 of TMZ o , cell proliferation p , colony ability q , apoptosis r , migration s and t and invasion u were measured by CCK-8, colony formation, flow cytometry, wound healing and transwell analyses respectively in resistant cells transfected with sh-NC + anti-NC, sh-circ_0072083 + anti-NC or sh-circ_0072083 + anti-miR-1252-5p after treatment of the indicated concentration of TMZ. * P < 0.05

Article Snippet: For transwell migration analysis, 24-well transwell chambers (Corning Inc., Corning, NY, USA) with a fibronectin-coated polycarbonate membrane were used.

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Luciferase, Activity Assay, Transfection, Construct, Western Blot, Migration, CCK-8 Assay, Flow Cytometry, Concentration Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Warburg effect-promoted exosomal circ_0072083 releasing up-regulates NANGO expression through multiple pathways and enhances temozolomide resistance in glioma

doi: 10.1186/s13046-021-01942-6

Figure Lengend Snippet: Exosomal circ_0072083 from resistant cells increases the resistance of sensitive glioma cells. a Exosomal circ_0072083 level was detected by qRT-PCR in resistance cells transfected with sh-circ_0072083 or sh-NC. b and c circ_0072083, miR-1252-5p, ALKBH5 and NANOG levels were examined in sensitive cells after treatment of exosome from resistant cells or PBS. IC50 of TMZ d , cell proliferation e and f , colony ability g , apoptosis h , migration i and j and invasion k were measured by CCK-8, colony formation, flow cytometry, wound healing and transwell analyses respectively in sensitive cells with stimulation of exosome from resistant cells or PBS after treatment of the indicated concentration of TMZ. * P < 0.05

Article Snippet: For transwell migration analysis, 24-well transwell chambers (Corning Inc., Corning, NY, USA) with a fibronectin-coated polycarbonate membrane were used.

Techniques: Quantitative RT-PCR, Transfection, Migration, CCK-8 Assay, Flow Cytometry, Concentration Assay