Qing Cong1, Bin Li1, Yisheng Wang1, Wenbi Zhang1, Mingjun Cheng1, Zhiyong Wu1, Xiaoyan Zhang1, Wei Jiang1, Congjian Xu2. 1. Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China. 2. Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China ; Department of Obstetrics and Gynecology of Shanghai Medical School, Fudan University Shanghai, P.R. China ; Institute of Biomedical Sciences, Fudan University Shanghai, P.R. China.
Abstract
OBJECTIVE: Mouse bone marrow mesenchymal stem cells (BMSCs) have been demonstrated to differentiate into female endometrial epithelial cells (EECs) in vivo. Our previous studies demonstrated that BMSCs can differentiate in the direction of EECs when co-cultured with endometrial stromal cells in vitro. Here, we obtain and analyse differential proteins and their relevant pathways in the process of BMSCs differentiating into EECs by isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis. METHODS: A 0.4-μm pore size indirect co-culture system was established with female mice endometrial stromal cells (EStCs) restricted in the upper Transwell chamber and BMSCs in the lower well plate. After indirect co-culture for several days, the BMSCs were revealed to progressively differentiate towards EECs in vitro. Then, four groups were divided according to different co-culture days with single culture groups of BMSCs as controls. Proteins were detected using iTRAQ based on 2DLC-ESI-MS/MS and data were analysed by bioinformatics. RESULTS: A total number of 311 proteins were detected, of which 210 proteins were identified with relative quantitation. Among them, 107 proteins were differentially expressed with a 1.2-fold change as the benchmark, with 61 up-regulated and 46 down-regulated proteins. Differential proteins CK19 and CK8 were epithelial markers and upregulated. Stromal marker vimentin were downregulated. Top canonical pathways was "remodeling of epithelial adhesions junctions" and "actin cytoskeleton signaling". Top networks was "cell-to-cell signaling and interaction, tissue development and cellular movement" regulated by ERK/MAPK and α-catenin. CONCLUSION: To the best of our knowledge, this is the first preliminary study of differential protein expression in the differentiation process of BMSCs into EECs in vitro. We further elucidated BMSCs differentiated in the direction of EECs. In addition, ERK/MAPK and α-catenin played important roles by regulating core differential proteins in the "cell-to-cell signaling and interaction, tissue development and cellular movement" network.
OBJECTIVE:Mouse bone marrow mesenchymal stem cells (BMSCs) have been demonstrated to differentiate into female endometrial epithelial cells (EECs) in vivo. Our previous studies demonstrated that BMSCs can differentiate in the direction of EECs when co-cultured with endometrial stromal cells in vitro. Here, we obtain and analyse differential proteins and their relevant pathways in the process of BMSCs differentiating into EECs by isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis. METHODS: A 0.4-μm pore size indirect co-culture system was established with female mice endometrial stromal cells (EStCs) restricted in the upper Transwell chamber and BMSCs in the lower well plate. After indirect co-culture for several days, the BMSCs were revealed to progressively differentiate towards EECs in vitro. Then, four groups were divided according to different co-culture days with single culture groups of BMSCs as controls. Proteins were detected using iTRAQ based on 2DLC-ESI-MS/MS and data were analysed by bioinformatics. RESULTS: A total number of 311 proteins were detected, of which 210 proteins were identified with relative quantitation. Among them, 107 proteins were differentially expressed with a 1.2-fold change as the benchmark, with 61 up-regulated and 46 down-regulated proteins. Differential proteins CK19 and CK8 were epithelial markers and upregulated. Stromal marker vimentin were downregulated. Top canonical pathways was "remodeling of epithelial adhesions junctions" and "actin cytoskeleton signaling". Top networks was "cell-to-cell signaling and interaction, tissue development and cellular movement" regulated by ERK/MAPK and α-catenin. CONCLUSION: To the best of our knowledge, this is the first preliminary study of differential protein expression in the differentiation process of BMSCs into EECs in vitro. We further elucidated BMSCs differentiated in the direction of EECs. In addition, ERK/MAPK and α-catenin played important roles by regulating core differential proteins in the "cell-to-cell signaling and interaction, tissue development and cellular movement" network.
Authors: Philip L Ross; Yulin N Huang; Jason N Marchese; Brian Williamson; Kenneth Parker; Stephen Hattan; Nikita Khainovski; Sasi Pillai; Subhakar Dey; Scott Daniels; Subhasish Purkayastha; Peter Juhasz; Stephen Martin; Michael Bartlet-Jones; Feng He; Allan Jacobson; Darryl J Pappin Journal: Mol Cell Proteomics Date: 2004-09-22 Impact factor: 5.911
Authors: Louie Ye; Robyn Mayberry; Camden Y Lo; Kara L Britt; Edouard G Stanley; Andrew G Elefanty; Caroline E Gargett Journal: PLoS One Date: 2011-06-15 Impact factor: 3.240