摘要
目的研究已建立的放射抗拒细胞对比模型基因表达谱差异,初步分析放射抗拒的相关基因。方法Hep-2细胞重复照射12次后,传代培养至细胞形态、增殖状态稳定,筛选得到抗拒细胞株Hep-2R。从亲代Hep-2细胞和Hep-2R细胞提取总RNA,选用2张各含基因14000个的基因表达谱cDNAV芯片检测。筛选2张芯片差异相同的基因,计算Hep-2R细胞与Hep-2细胞基因表达的比值。结果Hep-2R与Hep-2的放射生物学参数分别为SF2=0.6798,D0=3.24、α=0.2951、β=0.0363,及SF2=0.4148、Do=2.06、α=0.1074、β=0.0405,Hep-2R获得了明显的抗拒性(SF2的x^2=63.957,P〈0.001)。Hep-2R细胞中上调基因22个,下调基因19个,主要基因类别为:蛋白质合成基因11个、DNA损伤修复基因9个、细胞结构基因6个、凋亡相关基因3个、癌基因与抑癌基因2个、细胞周期基因2个、代谢基因2个、信号通路受体基因2个、其他类基因4个。端粒保护蛋白基因POT1表达在所有上调基因中变化最显著(上调3.348倍)。结论不同放射敏感性的对比模型细胞基因表达谱不同,POT1基因变化最显著,有可能成为放射增敏的靶点。
Objective To study the difference of gene expressions by the contrastive model including the cells with same pathological origin and genetic background, but definitely different radiorespense, and to find the main molecular targets related to radiosensitivity. Methods Human larynx squamous carcinoma cell, Hep -2 was irradiated with dose of 637 cGy repeatedly to establish a radioresistant daughter cell line. The radiobiology characteristics were obtained using clone forming assay. The difference of gene expression between parent and daughter cells was detected by cDNA microarray using two different arrays including 14 000 genes respectively. Results A radioresistant cell strain Hep -2R was isolated from its parental strain Hep -2 cell. The SF2 ,Do , α ,β for Hep - 2R cell line were 0. 6798,3.24,0. 2951 and 0. 0363, respectively, while 0. 4148,2. 06 ,0. 1074 and 0. 0405 for Hep - 2, respectively ( for SF2 ,x^2 = 63.957, P 〈 0. 001 ). Compared with Hep- 2 cells,the expressions of 41 genes were significantly altered in the radioresistant Hep- 2R cells, including 22 genes up - regulated and 19 genes down - regulated, which were involved in DNA repair, regulation of the cell cycle, cell proliferation, cytoskeleton, protein synthesis, cellular metabolism and especially apeptosis which is responsible for the different radiosensitivity between these two larynx cancer cells. The telomere protection protein gene, POT1 ,was the mostly up - regulated by 3. 348 times. Conclusions There is difference of gene expression between the radioresistant contrastive models. POT1 gene may be the target of radiosensitization.
出处
《中华放射医学与防护杂志》
CAS
CSCD
北大核心
2007年第6期526-529,共4页
Chinese Journal of Radiological Medicine and Protection
基金
湖北省自然科学基金资助项目(2004ABA165,2006ABA253)
关键词
放射敏感性
基因表达谱
端粒保护蛋白基因POT1
Radioresistance
DNA microarray expression
Telomere protection protein gene( POT1 )