Comparative profiling of miRNA expression of lung adenocarcinoma cells in two-dimensional and three-dimensional cultures |
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Authors: | Cui Li Hong T. Nguyen Yan Zhuang Zhen Lin Erik K. Flemington Ying Zhuo Stephen P. Kantrow Gilbert F. Morris Deborah E. Sullivan Bin Shan |
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Affiliation: | 1. Xiangya Hospital, Central South University, Hunan, China;2. Department of Medicine, Tulane School of Medicine, New Orleans, LA 70112, USA;3. Department of Medicine, Louisiana State University Health Sciences Center, 1901 Perdido Street, New Orleans, LA 70112, USA;4. Department of Pathology, Tulane School of Medicine, New Orleans, LA 70112, USA;5. Department of Microbiology and Immunology, Tulane School of Medicine, New Orleans, LA 70112, USA |
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Abstract: | Three-dimensional organotypic culture using reconstituted basement membrane matrix (rBM 3-D) is an invaluable tool to characterize morphogenesis of epithelial cells and to elucidate the tumor-modulating actions of extracellular matrix. microRNAs (miRNA) are a novel class of tumor modulating genes. A substantial amount of investigation of miRNAs in cancer is carried out using monolayer 2-D culture on plastic substratum, which lacks a consideration of the matrix-mediated regulation of miRNAs. In the current study we compared the expression of miRNAs in rBM 3-D and 2-D cultures of two lung adenocarcinoma cell lines. Our findings revealed a profound difference in miRNA profiles between 2-D and rBM 3-D cultures of lung adenocarcinoma cells. The rBM 3-D culture-specific miRNA profile was highlighted with higher expression of the tumor suppressive miRNAs (i.e., miR-200 family) and lower expression of the oncogenic miRNAs (i.e., miR-17–92 cluster and miR-21) than that of 2-D culture. Moreover, the expression pattern of miR-17, miR-21, and miR-200a in rBM 3-D culture correlated with the expression of their targets and acinar morphogenesis, a differentiation behavior of lung epithelial cells in rBM 3-D culture. Over-expression of miR-21 suppressed its target PTEN and disrupted acinar morphogenesis. In summary, we provide the first miRNA profile of lung adenocarcinoma cells in rBM 3-D culture with respect to acinar morphogenesis. These results indicate that rBM 3-D culture is essential to a comprehensive understanding of the miRNA biology in lung epithelial cells pertinent to lung adenocarcinoma. |
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Keywords: | ECM, extracellular matrix BM, basement membrane matrix rBM 3-D, reconstituted basement membrane matrix based three-dimensional organotypic culture miRNA, microRNA F-actin, filamentous actin qRT-PCR, quantitative RT-PCR pri-miR-21, primary miR-21 VMP1, vacuole membrane protein 1 3&prime UTR, 3&prime untranslated region ZEB2, Zinc finger E-box-binding homeobox 2 PTEN, phosphatase and tensin homolog |
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