Journal: Cell Death & Disease

Article Title: Pleiotropic actions of miR-21 highlight the critical role of deregulated stromal microRNAs during colorectal cancer progression

doi: 10.1038/cddis.2013.213

Figure Lengend Snippet: Ectopic miR-21 expression in MRC5 fibroblasts leads to upregulated MMP2 activity – SW480 CRC cell invasion is MMP2 dependent. ( a ) Representative experiment showing zymographic activity in CM 21 or CM SCC , with and without MMP2 inhibitor (MMP2inh). ( b ) Assessment of MMP2 activity by determination of zymographical band intensity. Results are expressed as a ratio of MMP2 activity in CM 21 compared with CM SCC in three independent experiments. ( c ) Transwell invasion assay using CM 21 or CM SCC as chemoattractant, with and without MMP2 inhibitor. Results represent the mean number of invading cells from four repeat experiments (* P <0.05)

Article Snippet: Specific MMP2 inhibitor I (Merck, Hoddesdon, UK), prepared at a stock concentration of 10 mM in DMSO was also added to the chemoattractant at a final concentration of 25 μ M, which achieved maximal inhibition of invasion with no cytotoxic effects (data not shown).

Techniques: Expressing, Activity Assay, Transwell Invasion Assay

Journal: Cell Death & Disease

Article Title: Pleiotropic actions of miR-21 highlight the critical role of deregulated stromal microRNAs during colorectal cancer progression

doi: 10.1038/cddis.2013.213

Figure Lengend Snippet: Proposed impact of deregulated miR-21 expression in CRC-associated stroma. Reciprocal signalling between stromal and epithelial cells (thin black arrows; left panel) is essential for normal colonic tissue homoeostasis. Early in cancer, this process is corrupted and secreted factors from transformed epithelial cells (dotted black arrow; central panel) initiate profound changes in adjacent stromal cells. Upregulated miR-21 expression in stromal fibroblasts supports TGF- β -dependent fibroblast-to-myofibroblast transdifferentiation (thick black arrow) and causes downregulated RECK expression. Myofibroblast-derived factors are important mediators of tumour progression (dotted red arrow; right panel), and although in the absence of TGF- β , miR-21 is not sufficient to initiate myofibroblast transdifferentiation, upregulated miR-21 does promote tumour proliferation and chemoresistance and enhanced tumour invasion through upregulated MMP2 activity

Article Snippet: Specific MMP2 inhibitor I (Merck, Hoddesdon, UK), prepared at a stock concentration of 10 mM in DMSO was also added to the chemoattractant at a final concentration of 25 μ M, which achieved maximal inhibition of invasion with no cytotoxic effects (data not shown).

Techniques: Expressing, Transformation Assay, Derivative Assay, Activity Assay

Journal: Oncotarget

Article Title: Filamin C, a dysregulated protein in cancer revealed by label-free quantitative proteomic analyses of human gastric cancer cells

doi:

Figure Lengend Snippet: (A) DU145 cells with filamin C silencing were fluorescence-labeled and analyzed in a zebrafish cancer metastasis model. The selected pictures were shown from 0 to 3 days post injection. The areas indicated with arrows were enlarged for visualizing disseminated cancer cells. (B) Filamin C was silenced in DU145 cells and the proenzyme (Pro) and activated form of MMP2 were analyzed. (C) Migration of DU145 cells with or without filamin C silencing were analyzed. The third group with filamin C silencing was further treated with the MMP2 inhibitor ARP100. The statistical analysis results were shown in Figure and the Western blot results were shown in Figure . FLNC, filamin C.

Article Snippet: After cultured for 36 h, the cells transfected with filamin C siRNA were treated with 50 μM ARP100 MMP2 inhibitor (Apexbio, Shanghai, China) and cultured for 12 h before the evaluation of inhibitory effects.

Techniques: Fluorescence, Labeling, Injection, Migration, Western Blot