Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 3 WNT11 is a novel ligand for ROR2 in humans. a RNA-Seq of MCF-7 pROR2 cells: Network of differentially expressed genes associated with non-canonical WNT signaling grouped according to their cellular localization. b MCF-7 cells were stimulated for 24 h with rWNT5A (100 ng/ml) and WNT11 expression was analyzed by western Blot. c+d Expression of the non-canonical WNT ligands was measured by qRT-PCR in MCF-7 (c) or the indicated ROR2-overexpressing human breast cancer cell lines (d) (mean ± SD, n = 3–9, *p < 0.05, p < 0.01, n.e. = not expressed). Expression values were calculated relative to the empty vector control cells. e Co-immunoprecipitation (Co-IP) of V5-WNT11 in MCF-7 pROR2 cells detects ROR2 by western blot. f Schematic representation of the ROR2 N-terminal deletion constructs. g Co-IP of V5-Wnt11 in MCF-7 expressing either pROR2-FL or pROR2-ΔΔ. h Cell invasion assays of MCF-7 expressing N-terminal ROR2 deletion constructs (mean ± SD, n = 3, *p < 0.01, **p = 0.0001, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: RNA Sequencing, Expressing, Western Blot, Quantitative RT-PCR, Plasmid Preparation, Control, Immunoprecipitation, Co-Immunoprecipitation Assay, Construct, Comparison

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 4 WNT11 mediates the pro-tumoral effects of ROR2. a+b Invasion assay: MCF-7 pcDNA or pROR2 cells were treated with WNT inhibitors (a) or Porcupine inhibitors (b) (mean ± SD, n = 3, *p < 0.05, **p < 0.01, ***p < 0.0001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c Invasion assay of MCF-7 pROR2 cells stably expressing a non-sense control (ns ctl) or WNT11 (shWNT11) shRNA (mean ± SD, n = 3, *p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. d Invasion of MCF-7 pROR2 cells transfected with control (siCTL) or WNT11 siRNA (siWNT11) +/− rhWNT11 (100 ng/ml) was assessed in Boyden chambers (mean ± SD, n = 3, *p < 0.05, **p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. e Invasion assay of BT-474 pcDNA and pROR2 cells transfected with either siCTL or siWNT11 (mean ± SD, n = 3, *p < 0.05, **p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. f AFM of cell-cell-junctions in the indicated cell lines. g Immunofluorescence for the tight junction protein ZO-1. h ECIS measurements of MCF-7 cells (box: 25-75th percentile, line at median, *p < 0.0001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. i+j xCELLigence measurements of MCF-7 cells (mean ± SD). Shown is one representative example (i) and a corresponding Area Under the Curve (AUC) analysis for all three independent experiments (mean ± SD, *p < 0.01). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: Invasion Assay, Comparison, Stable Transfection, Expressing, Control, shRNA, Transfection, Immunofluorescence

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 5 WNT11 mediates ROR2 signaling. a+b Western blot: RHOA and ROCK2 in MCF-7 and BT-474 pROR2 cells treated with control siRNA (siCTL) or siRNA against WNT11 (siWNT11) (a) with corresponding densitometric quantification normalized on HSP90 expression (b) (mean ± SD, *p < 0.05, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c Levels of active RHOA-GTP in the indicated cells were assessed by ELISA (mean ± SD, *p < 0.05, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c MCF-7 pcDNA and pROR2 siCTL/siWNT11 cells were characterized by RPPA for phospho-proteins associated with the WNT signaling pathway (n = 3, *p < 0.05). d Schematic representation of the master regulator analysis of ROR2/WNT11 signaling. The illustration was created with BioRender.com

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: Western Blot, Control, Expressing, Comparison, Enzyme-linked Immunosorbent Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 6 ROR2/WNT11 are expressed in metastatic breast cancer and associated with poor survival. a Pathway (pw) enrichment for RNA-Seq data from 31 patients with brain metastases given for different WNT subpathways. Significance was calculated with a log rank test. b qRT-PCR: Expression of ROR2 and WNT11 in samples of human brain metastases (line at median). The upper numbers indicate the number of samples with positive signals out of all investigated samples. c Kaplan-Meier survival curves showing the OS of metastatic patients based on their averaged WNT11 and ROR2 expression. The separation high/low was computed based on an optimal cutoff using the maxstat method. Significance was calculated with a log rank test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: RNA Sequencing, Quantitative RT-PCR, Expressing

Journal: BMC oral health

Article Title: Coexpression network analysis identified MT3 as a hub gene that promotes the chemoresistance of oral cancer by regulating the expression of YAP1.

doi: 10.1186/s12903-023-03600-z

Figure Lengend Snippet: Fig. 5 MT3 promotes cisplatin resistance by modulating YAP activation in OSCC. (A-B) Western blotting showed that YAP1 expression was decreased after MT3 knockdown, while the expression of MT3 was not changed after YAP1 knockdown in CAL27-CISR or Fadu-CISR cells. (C-D) Immunohistochemical staining of YAP1 in chemotherapy-resistant OSCC tissue (CR) and chemotherapy-sensitive tissue (CS). p < 0.01**. (E) qRT‒PCR results showing YAP1 mRNA levels in CAL27-CISR or Fadu-CISR cells. (F) qRT‒PCR results showing downstream target genes of YAP1, BIRC2, CNNT1, and CTGF mRNA levels in CAL27- CISR cells. (G) BIRC2, CNNT1, and CTGF mRNA levels in Fadu-CISR cells. n = 3, p < 0.01**

Article Snippet: Fadu and CAL27 oral cancer cell lines were transfected with sgRNA against MT3 or YAP1 cloned in PX459 (Addgene, USA).

Techniques: Activation Assay, Western Blot, Expressing, Knockdown, Immunohistochemical staining, Staining

Journal: BMC oral health

Article Title: Coexpression network analysis identified MT3 as a hub gene that promotes the chemoresistance of oral cancer by regulating the expression of YAP1.

doi: 10.1186/s12903-023-03600-z

Figure Lengend Snippet: Fig. 6 YAP1 inhibition suppresses the development of CIS resistance. (A) Cell survival declined after verteporfin (VP) treatment in CAL27-CISR or Fadu- CISR cells. (B) Cell proliferation was suppressed after VP treatment in CAL27-CISR or Fadu-CISR cells for 24 h, 48 and 72 h. (C) Compared with the DMSO group, the number of colonies was reduced after VP treatment in CAL27-CISR or Fadu-CISR cells. The results are presented as the mean ± SEM of three independent experiments. (D-G) CAL27-CISR or Fadu-CISR cells were subcutaneously inoculated into nude mice (n = 6) and then treated with DMSO (negative control), cisplatin, verteporfin, or cisplatin combined with verteporfin. Tumor growth (D, F) and survival analysis (E, G) were determined as indicated. (p < 0.05*, p < 0.01**, p < 0.001***)

Article Snippet: Fadu and CAL27 oral cancer cell lines were transfected with sgRNA against MT3 or YAP1 cloned in PX459 (Addgene, USA).

Techniques: Inhibition, Negative Control