Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CIP2A mediates fibronectin-induced bladder cancer cell proliferation by stabilizing β-catenin

doi: 10.1186/s13046-017-0539-8

Figure Lengend Snippet: Stromal FN correlates positively with the levels of CIP2A and PCNA in bladder cancer tissues. a Representative photomicrographs of IHC staining for FN, CIP2A and PCNA in low FN group and high FN group were showed. b The labelling indexes of CIP2A and PCNA were assayed in low FN group and high FN group based on the IHC results (*** P < 0.001). c Western blotting analysis of FN and CIP2A expression level was performed in bladder cancer tissues ( n = 68). d Gray value of FN and CIP2A was measured by ImageJ software according to the western blotting results, and scatter plot with regression line showed a correlation of them using the Spearman’s correlation analysis. e and f According to the results of IHC, the expression of FN, CIP2A, and PCNA in NMIBC group and MIBC group were assayed (*** P < 0.001)

Article Snippet: CIP2A short hairpin RNA (sh-CIP2A) plasmid and CIP2A overexpressed plasmid (CIP2A-plasmid) which were ligated in pGV101 vector and pGV142 vector respectively, were purchased from Shanghai GeneChem Technologies Co., Ltd. (Shanghai, China).

Techniques: Immunohistochemistry, Western Blot, Expressing, Software

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CIP2A mediates fibronectin-induced bladder cancer cell proliferation by stabilizing β-catenin

doi: 10.1186/s13046-017-0539-8

Figure Lengend Snippet: FN induces cell proliferation and CIP2A expression in bladder cancer cells. a and b Cell viability of T24 and J82 cells incubated with FN (0, 5, 10 and 20 μg/mL) for 4 days was evaluated by using MTT assay (** P < 0.01, *** P < 0.001). c and d Cell cycle distribution of T24 and J82 cells treated with FN (0 and 20 μg/mL) for 48 h was evaluated by flow cytometry. The percentage of cells in each phase were shown (* P < 0.05, ** P < 0.01, *** P < 0.001). e and f CIP2A mRNA and protein expression levels of T24 and J82 cells were measured after FN treatment (0, 5, 10 and 20 μg/mL) for 48 h by qRT-PCR and western blotting respectively (** P < 0.01, *** P < 0.001)

Article Snippet: CIP2A short hairpin RNA (sh-CIP2A) plasmid and CIP2A overexpressed plasmid (CIP2A-plasmid) which were ligated in pGV101 vector and pGV142 vector respectively, were purchased from Shanghai GeneChem Technologies Co., Ltd. (Shanghai, China).

Techniques: Expressing, Incubation, MTT Assay, Flow Cytometry, Quantitative RT-PCR, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CIP2A mediates fibronectin-induced bladder cancer cell proliferation by stabilizing β-catenin

doi: 10.1186/s13046-017-0539-8

Figure Lengend Snippet: CIP2A mediates FN-induced bladder cancer cell proliferation. a CIP2A expression levels were examined in sh-CIP2A and sh-Control bladder cancer cells (T24 and J82). b sh-CIP2A and sh-Control bladder cancer cells (T24 and J82) were incubated with FN (0 and 20 μg/mL) for 2 weeks and allowed to form colonies. c and d The viability of sh-CIP2A and sh-Control bladder cancer cells (T24 and J82) incubated with FN (0 and 20 μg/mL) for 4 days was evaluated by using MTT assay (*** P < 0.001). e and f Cell cycle distribution of sh-CIP2A and sh-Control bladder cancer cells (T24 and J82) treated with FN (0 and 20 μg/mL) for 48 h was evaluated by flow cytometry. The percentage of cells in each phase were shown (* P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant)

Article Snippet: CIP2A short hairpin RNA (sh-CIP2A) plasmid and CIP2A overexpressed plasmid (CIP2A-plasmid) which were ligated in pGV101 vector and pGV142 vector respectively, were purchased from Shanghai GeneChem Technologies Co., Ltd. (Shanghai, China).

Techniques: Expressing, Control, Incubation, MTT Assay, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CIP2A mediates fibronectin-induced bladder cancer cell proliferation by stabilizing β-catenin

doi: 10.1186/s13046-017-0539-8

Figure Lengend Snippet: CIP2A is associated with β-catenin. a β-catenin mRNA levels were examined in sh-CIP2A and sh-Control bladder cancer cells (T24 and J82) (n.s., not significant). b CIP2A, β-catenin, c-myc, and CyclinD1 expression levels in sh-CIP2A and sh-Control bladder cancer cells (T24 and J82) were examined by western blotting. c and d sh-CIP2A and sh-Control bladder cancer cells (T24 and J82) were treated with CHX (100 μg/mL). At the indicated times (0, 1, 2, 3, and 4 h) after CHX treatment, cells were analyzed by western blotting. e and f The gray value of β-catenin were quantified using ImageJ software based on the western blotting results (* P < 0.05, ** P < 0.01). g Immunofluorescence staining analysis showed the colocalization of CIP2A and β-catenin in T24 and J82 cells. h and i Co-IP analysis showed the interaction between CIP2A and β-catenin in T24 and J82 cells. CIP2A and β-catenin were immunoprecipitated using antibody against β-catenin. IgG was used as a negative control

Article Snippet: CIP2A short hairpin RNA (sh-CIP2A) plasmid and CIP2A overexpressed plasmid (CIP2A-plasmid) which were ligated in pGV101 vector and pGV142 vector respectively, were purchased from Shanghai GeneChem Technologies Co., Ltd. (Shanghai, China).

Techniques: Control, Expressing, Western Blot, Software, Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Immunoprecipitation, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CIP2A mediates fibronectin-induced bladder cancer cell proliferation by stabilizing β-catenin

doi: 10.1186/s13046-017-0539-8

Figure Lengend Snippet: CIP2A mediates FN-induced bladder cancer cell proliferation in vivo. a Tumor growth curve of sh-CIP2A or sh-Control T24 cells’ bladder cancer subcutaneous xenograft tumors was built after FN or vehicle treatment for 5 weeks (* P < 0.05, ** P < 0.01). b Photographs of dissected xenograft tumors from nude mice after sacrificed were presented. c The weight of dissected xenograft tumors in each group was assayed (* P < 0.05). d The IHC staining was performed to detect the expression levels of FN, CIP2A, and PCNA in harvested tumor tissues, and representative photomicrographs of 400-fold high power fields in each group were showed

Article Snippet: CIP2A short hairpin RNA (sh-CIP2A) plasmid and CIP2A overexpressed plasmid (CIP2A-plasmid) which were ligated in pGV101 vector and pGV142 vector respectively, were purchased from Shanghai GeneChem Technologies Co., Ltd. (Shanghai, China).

Techniques: In Vivo, Control, Immunohistochemistry, Expressing