[Effects of E2F-1 overexpression on apoptosis of gastric cancer cells and expressions of apoptosis-related genes].

Lin-Hai Yan, Lei Li, Yu-Bo Xie, Qiang Xiao, Chang-Qing Wang
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引用次数: 4

Abstract

Background and objective: E2F transcription factor 1 (E2F-1) is an important transcription factor in cell cycle. This study was to investigate the effects of E2F-1 overexpression on apoptosis of gastric cancer MGC-803 cells and expressions of the downstream genes.

Methods: The apoptotic rates were measured by flow cytometry in MGC-803/E2F-1 cells, MGC-803/EV cells or untransfected MGC-803 cells. The total RNA was extracted from MGC-803/E2F-1 cells or MGC-803 cells, and cDNA was obtained by RT-PCR. Fluorescent (fluorescence exchange clip) probes marked by Cy5 and Cy3 were hybridized with gene chips containing 21522 human genes. Subsequently, the two signal images were scanned by Lux Scan 10K/A dual pathways laser scanner and analyzed by LuxScan3.0 image analysis software. RT-PCR was used to verify the target genes.

Results: The apoptotic rate of MGC-803/E2F-1 cells [(8.40+/-0.91)%] was higher than that of MGC-803/EV [(4.53+/-0.61)%] and MGC-803 cells [(4.97+/-0.47)%]. Fifteen differentially expressed apoptosis-related genes were detected, 4 of which were up-expressed and 11 were down-expressed genes, and the same results were verified by RT-PCR.

Conclusion: Overexpression of E2F-1 accelerates apoptosis of gastric carcinoma MGC-803 cells, which may be related to the 15 differentially expressed genes.

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[E2F-1过表达对胃癌细胞凋亡及凋亡相关基因表达的影响]。
背景与目的:E2F转录因子1 (E2F-1)是细胞周期中重要的转录因子。本研究旨在探讨E2F-1过表达对胃癌MGC-803细胞凋亡及下游基因表达的影响。方法:采用流式细胞术检测MGC-803/E2F-1细胞、MGC-803/EV细胞和未转染的MGC-803细胞的凋亡率。分别从MGC-803/E2F-1细胞或MGC-803细胞中提取总RNA, RT-PCR获得cDNA。用Cy5和Cy3标记的荧光(荧光交换夹)探针与含有21522个人类基因的基因芯片杂交。随后,用LuxScan 10K/A双通道激光扫描仪扫描两幅信号图像,用LuxScan3.0图像分析软件进行分析。采用RT-PCR对目的基因进行验证。结果:MGC-803/E2F-1细胞的凋亡率[(8.40+/-0.91)%]高于MGC-803/EV细胞[(4.53+/-0.61)%]和MGC-803细胞[(4.97+/-0.47)%]。检测到15个差异表达的凋亡相关基因,其中4个为上调表达基因,11个为下调表达基因,RT-PCR验证结果相同。结论:E2F-1过表达加速胃癌MGC-803细胞凋亡,可能与15个差异表达基因有关。
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