Journal: Oncology Letters

Article Title: Expression of CD74 in bladder cancer and its suppression in association with cancer proliferation, invasion and angiogenesis in HT-1376 cells

doi: 10.3892/ol.2018.8309

Figure Lengend Snippet: Association between MIF and CD74 expression with clinical and pathological characteristics of patient samples.

Article Snippet: The CD74 shRNA lentiviral particles (cat. no., sc-35023-V; 1.0×10 6 /200 μl; Santa Cruz Biotechnology, Inc.) targeting the human CD74 transcript and control shRNA lentiviral particles (cat. no., sc-108080; 1.0×10 6 /200 μl; Santa Cruz Biotechnology, Inc.) were used to knock down CD74 in HT-1376 cells.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Expression of CD74 in bladder cancer and its suppression in association with cancer proliferation, invasion and angiogenesis in HT-1376 cells

doi: 10.3892/ol.2018.8309

Figure Lengend Snippet: Expression of MIF and CD74 in tissue samples and cells, and protein expression levels in HT-1376 cells following knockdown of CD74. (A) Little or no expression of immunoreactive CD74 was identified in the urothelial layers of normal bladder and NMIBC samples, but sections of MIBC samples exhibited strong immunoreactive signals of CD74. In contrast, MIF demonstrated strong immunoreactions in the normal and UCB samples. (B) Western blotting assay results for MIF and CD74 in the cultured urothelial SV-HUV-1, SW780, 5637, T24, J82 and HT-1376 cell lines. HT-1376 was the only one identified to express CD74, but all cells expressed MIF to a certain extent. (C) Immunofluorescence microscopy (×100, magnification) indicated positive CD74 staining in HT-1376 cells. (D) CD74 shRNA lentiviral particles abrogated the RNA expression of CD74 in the HT-1376 cells compared with untreated and scramble cells. (E) CD74 short hairpin RNA lentiviral particles abrogated the protein expression of CD74 in the HT-1376 cells when compared with the untreated and scramble cells. (F) Knockdown of CD74 modulated the expression levels of not only Cyclin D1, Cyclin E, but also intranuclear NF-κB p65, pAkt, pErk1/2. (G and H) However, no significant changes in Bcl-2, Bad, cleaved PARP and Erk1/2 were observed among all groups. *intranuclear NF-κB. NMIBC, non-muscle-invasive bladder cancer; MIBC, muscle-invasive bladder cancer; UCB, urothelial cell carcinoma of the bladder; MIF, macrophage migration inhibitory factor; CD74, cluster of differentiation; p65, transcription factor p65; Akt, RAC-alpha serine/threonine protein kinase; p, phosphorylated; Erk1/2, Extracellular regulated protein kinase 1/2; Bcl-2, B-cell lymphoma 2; Bcl-xL, Bcl-extra large; Bax, Bcl-2-associated X protein; PARP, poly(adenosine 5′-diphosphate-ribose) polymerase.

Article Snippet: The CD74 shRNA lentiviral particles (cat. no., sc-35023-V; 1.0×10 6 /200 μl; Santa Cruz Biotechnology, Inc.) targeting the human CD74 transcript and control shRNA lentiviral particles (cat. no., sc-108080; 1.0×10 6 /200 μl; Santa Cruz Biotechnology, Inc.) were used to knock down CD74 in HT-1376 cells.

Techniques: Expressing, Knockdown, Western Blot, Cell Culture, Immunofluorescence, Microscopy, Staining, shRNA, RNA Expression, Migration

Journal: Oncology Letters

Article Title: Expression of CD74 in bladder cancer and its suppression in association with cancer proliferation, invasion and angiogenesis in HT-1376 cells

doi: 10.3892/ol.2018.8309

Figure Lengend Snippet: In vitro analysis of the effects of CD74-knockdown cells. (A) Knockdown of CD74 attenuated cell proliferation in HT-1376 cells compared with the scramble group. *P<0.05 and **P<0.01 vs. Control shRNA. (B) Flow cytometry indicated that knockdown of CD74 significantly increased the proportion of G1 stage cells, decrease G2 stage and S stage ones, compared with scramble shRNA cells. (C) The cell invasion assay demonstrated that knockdown of CD74 significantly attenuated the invasion ability of HT-1376 cells, (D) and the cells of two groups were counted (mean ± SD) and the data of CD74 shRNA group were presented by a percentage of control shRNA group. (E and F) ELISA test indicated that the secretion of (E) VEGF and MMP-9 (F-a) was significantly reduced in CD74-knockdown-HT-1376 cells compared to the shRNA control, while not significantly reduced in MMP-2 (F-b). CD74, cluster of differentiation; VEGF, vascular endothelial growth factor; MMP, matrix metalloproteinase; sh, short hairpin.

Article Snippet: The CD74 shRNA lentiviral particles (cat. no., sc-35023-V; 1.0×10 6 /200 μl; Santa Cruz Biotechnology, Inc.) targeting the human CD74 transcript and control shRNA lentiviral particles (cat. no., sc-108080; 1.0×10 6 /200 μl; Santa Cruz Biotechnology, Inc.) were used to knock down CD74 in HT-1376 cells.

Techniques: In Vitro, Knockdown, Control, shRNA, Flow Cytometry, Invasion Assay, Enzyme-linked Immunosorbent Assay

Journal: Oncology Letters

Article Title: Expression of CD74 in bladder cancer and its suppression in association with cancer proliferation, invasion and angiogenesis in HT-1376 cells

doi: 10.3892/ol.2018.8309

Figure Lengend Snippet: In vivo analysis of the effects of CD74-knockdown cells. (A) Volume of tumors derived from CD74-knockdown cells was significantly decreased compared with those derived from control cells in vivo . (B) The average weight of the tumors was significantly decreased in in the CD74 shRNA group. (C) Untreated HT-1376 xenograft tumors exhibited strong CD74 signals by immunochemistry. (D) CD74 in the wild-type J82 tumor was indicated using immunostaining. *P<0.05 and **P<0.01 vs. Control shRNA. Magnification, ×100.

Article Snippet: The CD74 shRNA lentiviral particles (cat. no., sc-35023-V; 1.0×10 6 /200 μl; Santa Cruz Biotechnology, Inc.) targeting the human CD74 transcript and control shRNA lentiviral particles (cat. no., sc-108080; 1.0×10 6 /200 μl; Santa Cruz Biotechnology, Inc.) were used to knock down CD74 in HT-1376 cells.

Techniques: In Vivo, Knockdown, Derivative Assay, Control, shRNA, Immunostaining

Journal: Cancer Medicine

Article Title: SNGH16 regulates cell autophagy to promote Sorafenib Resistance through suppressing miR‐23b‐3p via sponging EGR1 in hepatocellular carcinoma

doi: 10.1002/cam4.3020

Figure Lengend Snippet: The lncRNA microarray data from Hep3B and Hep3B/So cells

Article Snippet: Human LncRNA Microaray V1.0 (Arraystar) was used to analyze the data of five replicate samples, calculate the p ‐value of the gene expression difference and statistical significance, and use the Fold Change (FC) ≥1.5 or ≤−1.5, P < .05 for t test as a criterion for differential gene screening.

Techniques: Microarray

Journal: Nature Communications

Article Title: circNDUFB2 inhibits non-small cell lung cancer progression via destabilizing IGF2BPs and activating anti-tumor immunity

doi: 10.1038/s41467-020-20527-z

Figure Lengend Snippet: a A heatmap and volcano plot show 109 differentially expressed circRNAs in three paired samples of NSCLC by Arraystar Human Circular RNA Microarray. Cut off is Log 2 (fold change) > 1 or <−1.0, P value < 0.05. b Analysis for RNA levels of circNDUFB2 in additional 52 paired samples of NSCLC (left). Expression proportions of circNDUFB2 in histogram and pie chart (right). Log 2 (T/N expression) value > 1 as significantly higher expression, which <−1 as significantly lower expression, and between −1 and 1 as no significant change. N nontumorous tissue, T tumorous tissue. Data are presented as mean ± s.d. P values are calculated by paired two-sided t -test. n = 52 biologically independent paired tissues of NSCLC. c The backsplice junction site of circNDUFB2 was identified by Sanger sequencing. d PCR analysis for circNDUFB2 and β-actin in cDNA and genomic DNA. RH random hexamers, OdT oligo(dT) 18 primers, gDNA genomic DNA. Two independent experiments were carried out with similar results. e Analysis for RNA levels of circNDUFB2 and NDUFB2 after treatment with RNase R. n = 3 biologically independent samples. f Analysis for RNA abundance of circNDUFB2 and NDUFB2 treated with Actinomycin D (2 μg/ml) at the indicated time point. n = 3 biologically independent samples. g RNA levels of circNDUFB2 , NDUFB2 , β-actin , and U1 in the nuclear and cytoplasmic fractions of A549 and H1650 cells. n = 3 biologically independent samples. h RNA FISH analysis for circNDUFB2 in A549 cells, scale bar = 15 μm. Two independent experiments were carried out with similar results. Data are presented as mean ± s.d in e – g . P values are calculated by unpaired two-sided t -test in a , e – g .

Article Snippet: The labeled cRNAs were hybridized onto the Arraystar Human Circular RNA Microarray V1.0 (6x7K, Arraystar), which contains 5396 probes specific for human circular RNAs backsplice junction region.

Techniques: Microarray, Expressing, Sequencing