Journal: Molecular Therapy. Nucleic Acids

Article Title: circPTEN suppresses colorectal cancer progression through regulating PTEN/AKT pathway

doi: 10.1016/j.omtn.2021.05.018

Figure Lengend Snippet: Upregulation of circPTEN restrains malignant behaviors of CRC cells (A) qRT-PCR analysis of circPTEN overexpression efficiency in RKO and SW620 cells. (B and C) Evaluation of proliferation of transfected cells via colony formation and EdU (bar value = 150 μm) assays is shown. (D) Measurement of the apoptosis of transfected cells through TUNEL (bar value = 150 μm) assay is shown. (E and F) Analysis of the migratory and invasive capabilities of transfected RKO and SW620 cells via wound healing (bar value = 100 μm) and transwell (bar value = 70 μm) assays is shown. (G) Western blot analysis of EMT-related proteins in different groups is shown. (H) IF (bar value = 20 μm) analysis of E-cadherin and N-cadherin in transfected cells is shown. ∗∗p < 0.01.

Article Snippet: Matrigel-coated transwell chamber (Corning, Corning, NY) was commercially acquired for cell invasion assay.

Techniques: Quantitative RT-PCR, Over Expression, Transfection, TUNEL Assay, Western Blot

Journal: Molecular Therapy. Nucleic Acids

Article Title: circPTEN suppresses colorectal cancer progression through regulating PTEN/AKT pathway

doi: 10.1016/j.omtn.2021.05.018

Figure Lengend Snippet: circPTEN mediates malignant behaviors of CRC cells via AKT signaling pathway (A) Western blot analysis of protein expression in cells with circPTEN upregulation. (B) The knockdown efficiency of sh-PTEN was evaluated via qRT-PCR and western blot. (C) Western blot analysis of protein expression after transfection with different plasmids is shown. (D and E) Evaluation of cell proliferation in different groups via colony formation and EdU (bar value = 150 μm) assays is shown. (F) TUNEL (bar value = 150 μm) assay was performed to measure cell apoptosis in different groups. (G and H) Analysis of the migration and invasion abilities via wound healing (bar value = 100 μm) and transwell (bar value = 70 μm) analyses is shown. (I and J) EMT markers in different groups were detected via western blot and IF (bar value = 20 μm). ∗∗p < 0.01.

Article Snippet: Matrigel-coated transwell chamber (Corning, Corning, NY) was commercially acquired for cell invasion assay.

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Transfection, TUNEL Assay, Migration

Journal: Cell Death & Disease

Article Title: MiRNA199a-3p suppresses tumor growth, migration, invasion and angiogenesis in hepatocellular carcinoma by targeting VEGFA, VEGFR1, VEGFR2, HGF and MMP2

doi: 10.1038/cddis.2017.123

Figure Lengend Snippet: miR-199a-3p inhibits migration and angiogenesis in ECs by regulating cross talk between CACs and ECs. Endothelial cell recruitment assay in Boyden chamber: HUVEC cells were co-cultured with HepG2 and SNU449 separately. premiR-199a-3p and control vector plasmids were transfected either in ( a ) HUVEC or ( b ) in HCC cell lines. Bar diagram represented percentage of migrated cells though 8 μ m filter. In vitro tube formation in ECs was performed after similar transfection in either ( c ) HUVEC or ( d ) HCC cell lines. HUVECs were cultured on matrigel-coated plate with CM of cancer cell lines. Bars represented percentage of cellular branches. All the experiments were repeated three times. * P <0.05 and ** P <0.01, respectively

Article Snippet: For invasion assay, Matrigel basement membrane matrix coated transwell chambers (BD Biosciences) were used.

Techniques: Migration, Cell Culture, Plasmid Preparation, Transfection, In Vitro

Journal: Cell Death & Disease

Article Title: MiRNA199a-3p suppresses tumor growth, migration, invasion and angiogenesis in hepatocellular carcinoma by targeting VEGFA, VEGFR1, VEGFR2, HGF and MMP2

doi: 10.1038/cddis.2017.123

Figure Lengend Snippet: miR-199a-3p inhibits migration and invasion of cancer cells by regulating the cross talk between cancer cells (HepG2 or SNU449) and hepatic stellate cells (LX2). ( a ) Migrated cancer cells in transwell plate upon co-culturing with either LX2 cells or mock media. LX2 cells were transfected with vector and premiR-199a-3p, and SNU449 cells were either ( b ) co-cultured with transfected LX2 or ( c ) grown on matrigel pre-coated plate with CM of transfected LX2 cells. Average number of SNU449 cells migrated and invaded was counted. * P <0.05 and ** P <0.01

Article Snippet: For invasion assay, Matrigel basement membrane matrix coated transwell chambers (BD Biosciences) were used.

Techniques: Migration, Transfection, Plasmid Preparation, Cell Culture

Journal: PLoS ONE

Article Title: 14-3-3β Promotes Migration and Invasion of Human Hepatocellular Carcinoma Cells by Modulating Expression of MMP2 and MMP9 through PI3K/Akt/NF-κB Pathway

doi: 10.1371/journal.pone.0146070

Figure Lengend Snippet: (A) Matrigel-free Transwell assay was conducted to assess the migration ability of SMCC-7721 and Hep3B cells that were transfected with 14-3-3β expression construct or the empty vector pcDNA3.1. (B) Matrigel-based Transwell assay was performed to examine the invasiveness of SMCC-7721 and Hep3B with exogenous expression of 14-3-3β. (C) Matrigel-free migration assay and (D) Matrigel-based invasion assay were performed in CSQT-2 cells with and without expression of 14-3-3β-specific siRNA. This figure shows the representative images and the statistical analysis of three independent experiments. Values are expressed as the mean ± standard deviation. *** p <0.001.

Article Snippet: Cell migration ability was assessed by the Transwell assay with uncoated upper chambers (Corning, New York, USA) [ ], whereas cell invasion was examined by the Transwell assay with the upper chamber pre-coated with phenol red-free Matrigel (BD Biosciences, San Jose, CA, USA).

Techniques: Transwell Assay, Migration, Transfection, Expressing, Construct, Plasmid Preparation, Invasion Assay, Standard Deviation

Journal: PLoS ONE

Article Title: 14-3-3β Promotes Migration and Invasion of Human Hepatocellular Carcinoma Cells by Modulating Expression of MMP2 and MMP9 through PI3K/Akt/NF-κB Pathway

doi: 10.1371/journal.pone.0146070

Figure Lengend Snippet: (A) Expression of 14-3-3β, MMP2 and MMP9 in 14-3-3β-transfected SMCC-7721 and Hep3B cells. (B) Western blot analysis of phosphorylation status of various signaling proteins and the level of nuclear p65 in 14-3-3β-overexpressing HCC cells. β-actin was used as the internal control for total or cytosolic proteins, while Lamin A served as the internal control for nuclear proteins. The 14-3-3β-transfected Hep3B cells were treated with (C) LY294002 (20 μM), the inhibitor of PI3K/Akt signaling, or (D) PDTC (10 μM), the inhibitor of NF-κB signaling, for 5 h, followed by western blot analysis of the expression of MMP2 and MMP9. (E) The invasiveness of 14-3-3β-overexpressing cells with and without LY294002 (20 μM) or PDTC (10 μM) treatment was assessed by Matrigel-based Transwell assay. Values are expressed as the mean ± standard deviation of three independent experiments. *** p <0.001. (F) Western blot analysis was performed to determine the activation status of p-Akt and NF-κB in 14-3-3β-overexpressing cells after treating the cells with LY294002 (20 μM) for 5 h.

Article Snippet: Cell migration ability was assessed by the Transwell assay with uncoated upper chambers (Corning, New York, USA) [ ], whereas cell invasion was examined by the Transwell assay with the upper chamber pre-coated with phenol red-free Matrigel (BD Biosciences, San Jose, CA, USA).

Techniques: Expressing, Transfection, Western Blot, Transwell Assay, Standard Deviation, Activation Assay