胰腺癌细胞株基因组纯合子缺失的检测

Genomic analysis of homozygous deletion in pancreatic cancer cell lines by gene chip

目的通过基因芯片筛查，以期发现胰腺癌新的纯合子缺失基因。方法应用Affymetrix公司的GeneChip^R Human Mapping 100K Mapping microarray芯片，检测AsPC-1、BxPC-S等共21株人类胰腺癌细胞株的纯合子缺失区域，筛选这些区域中可能与胰腺癌发生和发展有关的基因，用PCR扩增方法验证。结果21株胰腺癌细胞共计发现60个纯合子缺失区域，以9p21．S出现的频率最高，达47．6％（10／21）。其中25个区域无已知基因，其余区域含有1～4个候选基因。选择26个可能与胰腺癌发生发展有关的基因，行42个PCR反应，35个反应无条带出现，证实为纯合性缺失，芯片的准确度为83．3％。结论应用高解析度的单核苷酸多态性基因芯片检测到胰腺癌新的纯合子缺失位点，为今后进一步发现新的胰腺癌抑癌基因提供线索。

Objecttive To scan for novel homozygous deletions in pancreatic cancer cell lines. Methods Genome-wide DNA homozygous deletions in 21 human pancreatic cancer cell lines were analyzed by using GeneChip？ Human Mapping 100K Mapping microarray, and the regions with candidate genes possibly related to the development of pancreatic cancer were verified by PCR. Results In the genome of 21 pancreatic cell lines, 60 regions were detected to be the homozygous deletion, the 9p21. 3 was the highest frequent homozygous deletion （ 10/21, 47.6% ）. 25 regions had no known gene; other regions had 1 to 4 candidate genes. 42 PCR reactions were done on 26 candidate genes possibly related to the development of pancreatic cancer and 35 reactions had no band, which meant the existence of homozygous deletion. The accuracy of the chip was 83.3%. Conclusions High-resolution single nucleotide polymorphism arrays were able to detect pancreatic cancer genome-wide homozygous deletion, and may identify novel candidate tumor suppression for future study.