Journal: Journal of Cellular and Molecular Medicine

Article Title: TP 53 INP 1 inhibits hypoxia‐induced vasculogenic mimicry formation via the ROS /snail signalling axis in breast cancer

doi: 10.1111/jcmm.13625

Figure Lengend Snippet: Relationship between TP53INP1 expression and clinicopathologic features, VM formation in breast cancer

Article Snippet: MCF‐7 cells were transfected with TP53INP1 shRNA (HSH022738‐LvRU6MP) or shRNA control (CSHCTR001‐LVRU6MP) purchased from GeneCopoeia, Inc.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: TP 53 INP 1 inhibits hypoxia‐induced vasculogenic mimicry formation via the ROS /snail signalling axis in breast cancer

doi: 10.1111/jcmm.13625

Figure Lengend Snippet: TP53INP1 expression in breast cancer tissues and pericarcinous tissues

Article Snippet: MCF‐7 cells were transfected with TP53INP1 shRNA (HSH022738‐LvRU6MP) or shRNA control (CSHCTR001‐LVRU6MP) purchased from GeneCopoeia, Inc.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: TP 53 INP 1 inhibits hypoxia‐induced vasculogenic mimicry formation via the ROS /snail signalling axis in breast cancer

doi: 10.1111/jcmm.13625

Figure Lengend Snippet: Difference of VE‐cadherin, Snail and HIF‐1α expression in TP53INP1‐positive and TP53INP1‐negative groups

Article Snippet: MCF‐7 cells were transfected with TP53INP1 shRNA (HSH022738‐LvRU6MP) or shRNA control (CSHCTR001‐LVRU6MP) purchased from GeneCopoeia, Inc.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells

doi: 10.3390/ijms161025450

Figure Lengend Snippet: Dose-dependent expression of TP53inp1 in immortalized human fibroblast cells (F11hT). Relative gene expression was measured by qPCR with the delta-delta cycle threshold (ΔΔ C t ) method as described in the Experimental Section. The data are derived from at least three independent experiments, and error bars show SEM of the mean. Gene expression in the F11hT cells is expressed in comparison with the sham-irradiated fibroblasts cells (calibrators), in which levels of expression are regarded as a level of one. Cells were harvested 2 h after γ-irradiation. One-way ANOVA was used for analysis. ( * p < 0.05, *** p < 0.001).

Article Snippet: According to the manufacturer’s protocol of Santa Cruz Biotechnology shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with three gene-specific shRNA expression lentiviral vectors (human TP53inp1 shRNA) transfected into subconfluent F11hT cells.

Techniques: Expressing, Derivative Assay, Irradiation

Journal: International Journal of Molecular Sciences

Article Title: TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells

doi: 10.3390/ijms161025450

Figure Lengend Snippet: TP53inp1 gene silencing in F11hT-NT and F11hT-shTP cells. ( A ) Values were calculated by qPCR with the ΔΔCT method. Data are given from at least four experiments, and error bars show SEM of the mean. Gene expression in the F11hT-shTP cells is compared with the sham-irradiated F11ht-NT cells, where the expression is fixed as a level of one. Statistical analysis was performed using one-way ANOVA-test ( * p < 0.05, *** p < 0.001). ( B ) Irradiation induces expression of TP53inp1 . TP53inp1 protein level was detected by Western blot at 24h post-irradiation with 2 and 6 Gy and normalized to Histone-H3. Expression of TP53inp1 protein was significantly lower in TP53inp1 silenced F11hT-shTP cells as compared to the F11hT-NT cells. Densitometric analysis of the bands, relative to Histone-H3, was performed using ImageJ softwer ( http://imagej.nih.gov/ij/ ).

Article Snippet: According to the manufacturer’s protocol of Santa Cruz Biotechnology shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with three gene-specific shRNA expression lentiviral vectors (human TP53inp1 shRNA) transfected into subconfluent F11hT cells.

Techniques: Expressing, Irradiation, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells

doi: 10.3390/ijms161025450

Figure Lengend Snippet: The effect of TP53inp1 silencing on the formation of radiation-induced autophagic vacuoles. ( A ) Quantitation of autophagic vacuoles shows a significant increase in the 6 Gy treated groups as compared to the sham irradiated F11hT-NT cells ( * p value < 0.5). Silencing of TP53inp1 is resulted significantly less autophagosome in 6 Gy-exposed F11hT-shTP cells ( * p value < 0.5). Two days after irradiation, cells were treated with Acridine Orange dye and red (autophagosome) puncta were counted from minimum eight cover slips ( n ≥ 8) under fluorescent microscope. White arrowheads denote the autophagic vacuoles. Results were analyzed with One-way ANOVA; ( B ) fluorescence photomicrograps obtained after Acridine Orange staining. Control cells (0 Gy) showing a few cytoplasmic AV formation, the number of AV increased in irradiated F11hT-NT cells and, to a lesser extent, in F11hT-shTP cells; ( C ) Representative flow cytometry plots are demonstrative of LC3B intracellular staining in response to 6 Gy exposures. Dot plot analysis is derived from the non-gated cell population. Flow cytometry analysis of F11hT-NT and F11hT-shTP cells using LC3B Antibody (Sigma, St. Louis, MI, USA) compared to a nonspecific isotype control antibody. Acquisition of 10,000 events was collected and for analysis the CellQuest software (BD Biosciences, San Jose, CA, USA) was used; ( D ) Representative flow cytometry histograms of percent LC3B-positive fibroblast are shown at right. Labeling of LC3B-positive cells at 48 h in F11hT-NT cells (right, top graph) and F11hT-shTP cells (right, bottom graph) after irradiation are graphed.

Article Snippet: According to the manufacturer’s protocol of Santa Cruz Biotechnology shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with three gene-specific shRNA expression lentiviral vectors (human TP53inp1 shRNA) transfected into subconfluent F11hT cells.

Techniques: Quantitation Assay, Irradiation, Microscopy, Fluorescence, Staining, Flow Cytometry, Derivative Assay, Software, Labeling

Journal: International Journal of Molecular Sciences

Article Title: TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells

doi: 10.3390/ijms161025450

Figure Lengend Snippet: Effect of TP53inp1 silencing on the accumulation of CD (common deletion) in the mitochondrial genome. Dose-dependent increase of mitochondrial common DNA deletions was compared in irradiated F11hT-NT and F11hT-shTP cells by qPCR. The mean ± SEM of at least three independent experiments are shown. Changes in the relative amount of CD were measured 72 h after the γ-irradiation. The mean ± SEM data derived from at least three experiments. Statistically significant changes calculated with One-way ANOVA are labeled as * p < 0.05.

Article Snippet: According to the manufacturer’s protocol of Santa Cruz Biotechnology shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with three gene-specific shRNA expression lentiviral vectors (human TP53inp1 shRNA) transfected into subconfluent F11hT cells.

Techniques: Irradiation, Derivative Assay, Labeling

Journal: International Journal of Molecular Sciences

Article Title: TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells

doi: 10.3390/ijms161025450

Figure Lengend Snippet: ( A ) Effect of TP53inp1 silencing on radiation induced senescence. Senescence associated-β-galactosidase positive F11hT-NT and F11hT-shTP cells was measured six days after exposure to a single dose of 6 Gy irradiation. Data presented are means ± SEM, n = 9 from three separate experiments. Statistical analysis was performed with one-way ANOVA followed by a Bonferroni post-test. A statistically significant difference p < 0.05 ( * ) is indicated; ( B ) Representative pictures of human fibroblasts (F11hT-NT) and TP53inp1 silenced fibroblasts (F11hT-shTP) were irradiated with 6Gy and stained with SA-βGal. The sham irradiated control shows exiguous staining, while the 6 Gy irradiated samples are powerfully stained. (Photo: Zeiss Axioskop2plus microscope, 100× magnification; Olympus Camedia camera; 3× optical zoom).

Article Snippet: According to the manufacturer’s protocol of Santa Cruz Biotechnology shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with three gene-specific shRNA expression lentiviral vectors (human TP53inp1 shRNA) transfected into subconfluent F11hT cells.

Techniques: Irradiation, Staining, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells

doi: 10.3390/ijms161025450

Figure Lengend Snippet: TP53inp1 silencing alters expression of IR–induced p53 targets. Graphs show relative transcript expression of CDKN1A in ( A ) panel GDF-15 and in ( B ) panel, as quantified by qPCR in F11hT-NT and F11hT-shTP fibroblasts without treatment and after 2 Gy γ ray exposure for 2 h ( * and ** are p < 0.05 and 0.01 compared with treated F11hT-NT and F11hT-shTP cells, respectively). Target transcript expression was normalized by the corresponding mean of housekeeping GAPDH and β-Actin values. Data are means of triplicates ± SEM. Statistically significant changes calculated with One-way ANOVA are labelled as * p < 0.05.

Article Snippet: According to the manufacturer’s protocol of Santa Cruz Biotechnology shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with three gene-specific shRNA expression lentiviral vectors (human TP53inp1 shRNA) transfected into subconfluent F11hT cells.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells

doi: 10.3390/ijms161025450

Figure Lengend Snippet: Oligonucleotide primers used in quantitative real time-PCR.

Article Snippet: According to the manufacturer’s protocol of Santa Cruz Biotechnology shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with three gene-specific shRNA expression lentiviral vectors (human TP53inp1 shRNA) transfected into subconfluent F11hT cells.

Techniques: