Li Hong-Zhen, Gao Xian-Shu, Xiong Wei, Zhao Jing, Zhang Hai, Zhou De-Min
Department of Radiation Oncology, Peking University First Hospital, Beijing 100034, PR China.
Chin J Cancer. 2010 Oct;29(10):882-8. doi: 10.5732/cjc.010.10148.
Radioresistant cells in esophageal cancer is one of the important reasons for the local failure of radiotherapy. In recent years, some researchers used gene chip technology to screen the differentially expressed genes between parental and radioresistant human esophageal cancer cells. But there were some problems in these studies, for example comparing cells at only one time interval, and genetic background not matching. In this study, we selected 3 different pairs of parental and radioresistant human esophageal cancer cells, and compared the gene expression profiles by cDNA microarray at 3 time intervals to identify and analyze the differentially expressed genes between parental and radioresistant human esophageal cancer cells.
We compared the gene expression profiles between parental cells (TE13, Seg-1, Kyse170) and radioresistant cells (TE13R, Seg-1R, Kyse170R) before, and at 8 h and 24 h after irradiation with a cDNA microarray consisting of 48 000 genes (Human Genome). We identified differentially expressed genes by Pathway and GO analyses, and verified the differentially expressed genes LEF1 and CTNNB1 by RT-PCR.
A total of 460, 451, and 397 differentially expressed genes were found before, and at 8 h and 24 h after irradiation. After Pathway and GO analyses, 14 differentially expressed genes, participating in cell growth, apoptosis, cell cycle regulation, gene repair and signal transmission, were selected to further research. LEF1 and CTNNB1 were verified by RT-PCR, and the results were consistent with those of cDNA microarray.
The WNT signal pathway may be an important pathway participating in the formation of radioresistance of esophageal cancer cells. LEF1 and CTNNB1 may be the important genes causing the esophageal cancer cell radioresistance.
食管癌中的放疗抵抗细胞是放疗局部失败的重要原因之一。近年来,一些研究者利用基因芯片技术筛选亲本和放疗抵抗的人食管癌细胞之间的差异表达基因。但这些研究存在一些问题,例如仅在一个时间间隔比较细胞,以及基因背景不匹配。在本研究中,我们选择了3对不同的亲本和放疗抵抗的人食管癌细胞,在3个时间间隔通过cDNA微阵列比较基因表达谱,以鉴定和分析亲本与放疗抵抗的人食管癌细胞之间的差异表达基因。
我们使用包含48000个基因(人类基因组)的cDNA微阵列,比较亲本细胞(TE13、Seg-1、Kyse170)和放疗抵抗细胞(TE13R、Seg-1R、Kyse170R)在照射前、照射后8小时和24小时的基因表达谱。我们通过通路和基因本体分析鉴定差异表达基因,并通过RT-PCR验证差异表达基因LEF1和CTNNB1。
在照射前、照射后8小时和24小时分别发现460、451和397个差异表达基因。经过通路和基因本体分析,选择了14个参与细胞生长、凋亡、细胞周期调控、基因修复和信号传导的差异表达基因进行进一步研究。通过RT-PCR验证了LEF1和CTNNB1,结果与cDNA微阵列一致。
WNT信号通路可能是参与食管癌细胞放疗抵抗形成的重要通路。LEF1和CTNNB1可能是导致食管癌细胞放疗抵抗的重要基因。