Identification of tumor invasion-related differentially expressed genes in different grades and all-trans retinoic acid-treated astrocytoma cell lines

作     者：Yi Zeng Zhong Yang Yangyun Han Chao You 

作者机构：Department of Neurosurgery West China Hospital Sichuan University Chengdu 610041 Sichuan Province China Department of Neurosurgery Deyang People＇s Hospital Deyang 618000 Sichuan Province China Department of Neurobiology Third Military Medical University of Chinese PLA Chongqing 400038 China 

出 版 物：《Neural Regeneration Research》 (中国神经再生研究（英文版）)

年 卷 期：2008年第3卷第11期

页      面：1222-1228页

核心收录：

学科分类：0710[理学-生物学] 1002[医学-临床医学] 1001[医学-基础医学(可授医学、理学学位)] 100214[医学-肿瘤学] 10[医学] 

基　　金：the Sichuan Provincial Bureau of Health No.050209 

主　　题：all-trans retinoic acid astrocytoma cDNA microarray CHG-5 invasion SHG-44 

摘      要：BACKGROUND： Although several genetic aberrations and gene expressional changes have been shown to exist in tumors and different grades of astrocytomas, as well as in normal tissues, the gene profiling and genetic pathways associated with malignant transformation and progression remain unclear. OBJECTIVE： To identify differentially expressed genes related to tumor invasion from various grades and all-trans retinoic acid （ATRA）-treated astrocytoma cell lines by cDNA microarray. DESIGN, TIME AND SETTING： In vitro gene experiment was performed at the Department of Neurobiology, Third Military Medical University of Chinese PLA from January to October 2007. MATERIALS： Two different grades of astrocytoma cell lines CHG-5 （WHO grade II ） and SHG-44 （WHO grade IV） were developed by our laboratory; a cell differentiation-inducing agent ATRA and a human cDNA microarray technology were used to determine differentially expressed genes （City University of Hong Kong）. METHODS： Total RNA was extracted using the Trizol test kit. Reverse transcription was performed using Superscript 11 reverse transcriptase. The cDNA product （target DNA） was marked with fluorochromes Cy3 （normal SHG-44） and Cy5 （CHG-5 or ATRA-treated SHG-44）, followed by chip hybridization. MAIN OUTCOME MEASURES： Gene expression profiles of CHG-5 vs. SHG-44 and ATRA-treated vs. normal SHG-44 were performed to identify differentially expressed genes. Several of these genes were randomly selected for Northern Blot analysis. The identification of genes that were similarly regulated （overlapping） was performed by comparing gene expression profiles between CHG-5 and SHG-44 cells, and between SHG-44 cells with or without treatment with ATRA. RESULTS： No significant differences were observed between CHG5 and SHG-44 cell line morphology. Under confocal microscopy, GFAP staining intensity of CHG5 cells was greater than SHG-44 cells （t = 6.078 P = 0.004）. Growth curve analysis demonstrated that the speed of SHG-44 cell growth