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Suppression of migratory/invasive ability and induction of apoptosis in adenomyosis- derived mesenchymal stem cells by cyclooxygenase-2 inhibitors  Yi-Jen.

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Presentation on theme: "Suppression of migratory/invasive ability and induction of apoptosis in adenomyosis- derived mesenchymal stem cells by cyclooxygenase-2 inhibitors  Yi-Jen."— Presentation transcript:

1 Suppression of migratory/invasive ability and induction of apoptosis in adenomyosis- derived mesenchymal stem cells by cyclooxygenase-2 inhibitors  Yi-Jen Chen, M.D., Hsin-Yang Li, M.D., Ph.D., Yuh-Lih Chang, Ph.D., Chiou-Chung Yuan, M.D., Lung-Kuo Tai, Ph.D., Kai Hsi Lu, Ph.D., Chia-Ming Chang, M.D., Shih-Hwa Chiou, M.D., Ph.D.  Fertility and Sterility  Volume 94, Issue 6, Pages e4 (November 2010) DOI: /j.fertnstert Copyright © Terms and Conditions

2 Figure 1 Isolation and characterization of endometrial mesenchymal stem-like cells (EMSCs) and adenomyosis-derived mesenchymal stem-like cells (AMSCs) from normal endometrium (EM) and adenomyosis. (A) Morphology of AMSCs (n = 6) and EMSCs (n = 9) isolated from tissue samples of 10 adenomyoses and 10 normal EM. Bar = 50 μm. (B) Flow cytometry analysis of AMSCs (positive: green line; control: purple). (C) Karyotype analysis of AMSCs. AMSCs at passage 10 show normal karyotypes of 46,XX. (D) Quantitative polymerase chain reaction analysis showed that mRNA expression of stemness-related genes in freshly isolated normal EM cells without adenomyosis, EMSCs, and AMSCs. (E) Multilineage differentiation capability of AMSCs. Under adipogenic conditions, AMSC-derived adipocytes were positively stained by oil red O assay. Under osteogenic conditions, AMSC-derived osteocytes were positively stained for mineralized matrices by alizarin red staining. Under myocytic differentiation, red indicates alpha-actinin stain. Under hepatogenic conditions, AMSC-derived hepatocyte-like cells were positive for periodic acid–Schiff assay. Data are the mean ± SD of three independent experiments. Fertility and Sterility  , e4DOI: ( /j.fertnstert ) Copyright © Terms and Conditions

3 Figure 2 Gene expression microarray analysis of endometrial mesenchymal stem-like cells (EMSCs) and adenomyosis-derived mesenchymal stem-like cells (AMSCs). (A) Unsupervised principal component analysis (PCA) of different samples. The genetic profiles of 3 AMSCs, 3 EMSCs, 3 bone marrow–derived mesenchymal stem cells (BMSCs), 3 embryonic stem cells (ESCs), and 10 normal endometrium (EM) samples were analyzed using PCA. The two-dimensional plot view of gene expression data is shown, with respect to their correlation with the first three principal components. (B) A heat map shows that 621 probe sets differentially expressed in 3 EMSCs and 3 AMSCs. Blue color means increased expression; red color means decreased expression. The significantly up- or down-regulated expression of genes (>2-fold or <0.5-fold, respectively) in AMSCs were compared with those in EMSCs. (C) Cyclooxygenase-2 (COX-2)–related genes in apoptosis pathway were predicated by Agilent literature pathway in Cytoscape. (D) COX-2–related genes in epithelial-mesenchymal transition (EMT) pathway were predicted by Pubgene ( Fertility and Sterility  , e4DOI: ( /j.fertnstert ) Copyright © Terms and Conditions

4 Figure 3 (A) Validation of inflammation-, apoptosis-, and epithelial-mesenchymal transition (EMT)–related genes using quantitative polymerase chain reaction. ∗The mRNA expression level is significantly higher in adenomyosis-derived mesenchymal stem-like cells (AMSCs) (P<.05). #The mRNA expression level is significantly higher in endometrial mesenchymal stem-like cells (EMSCs) (P<.05). (B) The cyclooxygenase-2 (COX-2) protein expression of AMSCs and EMSCs were verified by Western blot. COX-2 protein expression was decreased in AMSCs after treated with 40 μm COX-2 inhibitor (NS-398) for 48 hours. (C) AMSCs and EMSCs were treated with NS-398 for 24 and 48 hours. Cell viability was determined by cell number. (D) AMSCs and EMSCs were treated with 20 or 40 μm NS-398 for 48 hours. Cell apoptotic cells were determined by terminal deoxynucleotide transferase–mediated dUTP nick-end labeling assay. (E) Cell migration and invasive ability of AMSCs and EMSCs were analyzed. AMSCs showed decreased motility and invasiveness in both migration and invasion assays after treated with 20 or 40 μm NS-398 for 48 hours. Assays were performed for ≥3 times, and results are expressed as mean ± SD. ∗P<.05. Fertility and Sterility  , e4DOI: ( /j.fertnstert ) Copyright © Terms and Conditions

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