Journal: Archives of Medical Science : AMS

Article Title: MiR-1286 inhibits lung cancer growth through aerobic glycolysis by targeting PKM2

doi: 10.5114/aoms.2019.87812

Figure Lengend Snippet: MiR-1286 overexpression inhibited the proliferation of NSCLC cells. A – CCK8 assay was performed to detect A549 cell proliferation ability at indicated time points after transfection of miR-1286 mimic or control mimic and miR-1286 inhibitor or control inhibitor. B – Cell number was determined by cell counting after transfection of miR-1286 mimic or control mimic and miR-1286 inhibitor or control inhibitor for 48 h in A549 cells *P < 0.05, **p < 0.01.

Article Snippet: Cell Counting Kit-8 (CCK8) was bought from Transgen Biotech (Beijing, China).

Techniques: Over Expression, CCK-8 Assay, Transfection, Cell Counting

Journal: Archives of Medical Science : AMS

Article Title: MiR-1286 inhibits lung cancer growth through aerobic glycolysis by targeting PKM2

doi: 10.5114/aoms.2019.87812

Figure Lengend Snippet: MiR-1286 inhibited NSCLC cell proliferation via targeting PKM2. A – Cell proliferation was determined by CCK8 assay after transfection of miR-1286 mimic or control mimic, together with pcDNA3.1-PKM2 or pcDNA3.1 for 48 h. B – Cell proliferation was determined by CCK8 assay after transfection of miR-1286 inhibitor or control inhibitor, together with shPKM2 or sh-Scramble for 48 h. C – Lactate production was measured by using the Nova Bioprofile 100 analyzer after transfection miR-1286 mimic or control mimic, together with pcDNA3.1-PKM2 or pcDNA3.1 for 24 h. D – Lactate production was measured after transfection miR-1286 inhibitor or control inhibitor, together with shPKM2 or sh-Scramble for 48 h *P < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Cell Counting Kit-8 (CCK8) was bought from Transgen Biotech (Beijing, China).

Techniques: CCK-8 Assay, Transfection

Journal: Gland Surgery

Article Title: Tumor-derived mesenchymal progenitor cell-related genes in the regulation of breast cancer proliferation

doi: 10.21037/gs-23-387

Figure Lengend Snippet: Comparison of cell proliferation, migration, and secretion abilities of MSCs and MPCs. (A) The cell proliferation was measured via eFluor670 dye at 0, 24, 48, and 72 hours for MSCs and MPCs. (B) MSC and MPC proliferation indices of eFluor670 dye (n=5; *P<0.05, MSC vs. MPC). (C) MSC and MPC proliferation indices of CCK8 assay at 0, 24, 48, and 72 hours (n=5; ***P<0.001, MSC vs. MPC). (D) Cell scratch assay to detect the migration ability of MPCs and MSCs (n=5). (E) Detection of cellular inflammatory factors according to qRT-PCR (n=5; **P<0.01; ***P<0.001). MSC, mesenchymal stem cell; MPC, mesenchymal progenitor cell; IL-6, interleukin 6; CXCL12, C-X-C motif chemokine ligand 12; FGF2, fibroblast growth factor 2; TGF-β, transforming growth factor β; IGF-1, insulin-like growth factor 1; CCK8, Cell Counting Kit 8; qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: MSCs and MPCs were seeded into 96-well plates and processed at 0, 48, and 72 h. CCK8 (10 μL) reagent (ScienCell, Carlsbad, CA, USA) was added to each well, and the cell was cultured again for 2 h; the absorbance was measured at 450 nm with a microplate reader, the experimental results were recorded, and a growth curve was drawn.

Techniques: Comparison, Migration, CCK-8 Assay, Wound Healing Assay, Quantitative RT-PCR, Cell Counting, Real-time Polymerase Chain Reaction

Journal: Gland Surgery

Article Title: Tumor-derived mesenchymal progenitor cell-related genes in the regulation of breast cancer proliferation

doi: 10.21037/gs-23-387

Figure Lengend Snippet: STOM-overexpressing MPCs secrete inflammatory factors while promoting breast cancer proliferation. (A) Plasmid constructs mapping of LAMA5 , CCBE1 , and STOM . (B) qRT-PCR mRNA expression of target genes in MPCs transfected with overexpression plasmids (n=3; *P<0.05; ***P<0.001 target group compared with the control group). (C) qRT-PCR showed upregulation of CXCL12 and FGF2 expression after upregulation of the STOM gene (n=5; ***P<0.001 vs. control). (D) qRT-PCR showed upregulation of IGF expression after upregulation of the CCBE1 gene (n=5; ***P<0.001 vs. control). (E) qRT-PCR showed upregulation of FGF1 expression after upregulation of the LAMA5 gene (n=5; ***P<0.001 vs. control). (F-H) CCK8 assay indicated proliferation of MPCs overexpressing LAMA5 , CCBE1 , and STOM (n=5; ***P<0.001 vs. control). (I-K) CCK8 assay indicated proliferation of breast cancer MDA-MB-231 cells promoted by MPCs overexpressing STOM after coculture, and the overexpression of CCBE1 and LAMA5 did not have this pro-proliferative effect (n=5; ***P<0.001 vs. control). (L) CCK8 assay indicated proliferation of breast cancer MCF-7 cells promoted by MPCs overexpressing STOM after coculture (n=3; ***P<0.001 vs. control). MSC, mesenchymal stem cell; MPC, mesenchymal progenitor cell; qRT-PCR, quantitative real-time polymerase chain reaction; STOM, stomatin; CCBE1, collagen and calcium binding EGF domains 1; LAMA5, laminin subunit alpha 5; CXCL12, C-X-C motif chemokine ligand 12; FGF2, fibroblast growth factor 2; FGF1, fibroblast growth factor 1; IGF, insulin-like growth factor.

Article Snippet: MSCs and MPCs were seeded into 96-well plates and processed at 0, 48, and 72 h. CCK8 (10 μL) reagent (ScienCell, Carlsbad, CA, USA) was added to each well, and the cell was cultured again for 2 h; the absorbance was measured at 450 nm with a microplate reader, the experimental results were recorded, and a growth curve was drawn.

Techniques: Plasmid Preparation, Construct, Quantitative RT-PCR, Expressing, Transfection, Over Expression, Control, CCK-8 Assay, Real-time Polymerase Chain Reaction, Binding Assay