高转移卵巢上皮性癌基因表达谱中差异表达基因与染色体拷贝数变异的相关性

Study of the metastasis-associated genes and its copy numbers variation in highly metastatic epithelial ovarian cancer

目的采用基因芯片技术研究高转移卵巢上皮性癌（卵巢癌）细胞株HO-8910PM和正常卵巢上皮组织基因表达谱的差异表达基因及其与染色体拷贝数变异的相关性。方法利用全基因组表达谱芯片、单核苷酸多态性（SNP）扫描芯片分别检测高转移卵巢癌和正常卵巢上皮组织之间的差异表达基因以及染色体拷贝数变异情况，并采用生物信息学方法对检测结果进行相关性分析。筛选部分染色体片段和差异表达基因，用荧光原位杂交技术和实时荧光定量PCR技术进行验证。结果通过对表达谱芯片和SNP扫描芯片整合分析显示，高转移卵巢癌细胞中有379个SNP位点，共385个差异表达基因（其中有6个SNP位点发现各有2个基因）。这些SNP位点染色体拷贝数扩增的有240个，缺失的有139个。染色体定位分析发现，385个差异表达基因散在分布于各条染色体上，其中3号染色体最多，有34个（占8．8％，34／385）；其次是2、9、10、1和11号染色体分别有33个（8．6％）、28个（7．3％）、27个（7．0％）、25个（6．5％）、24个（6．2％）；6和12号染色体各有23个（6．0％）；4和5号染色体各有22个（5．7％）；这10条染色体占总数的67．8％（261／385）。基因的功能分类：385个差异表达基因属于酶及其调控子基因的最多（99个，占25．7％），其次是信号传导基因（54个，占14．0％），第3类是核酸结合基因（50个，占13．0％），第4类是蛋白结合基因（36个，占9．4％）；以上4大类共占差异表达基因总数的62．1％（239／385）。结论肿瘤的转移是多基因共同作用的结果。4大类（酶及其调控子、核酸结合、蛋白结合、信号传导）差异表达基因是今后研究卵巢癌转移相关的重要基因。1、2、3、4、5、6、9、11和12号染色体是值得关注的转移相关位点所在的染色体。

Objective To investigate the relationship of the metastasis-associated genes and its copy numbers variation in the highly metastatic human epithelial ovarian cancer cell line HO-8910PM. Methods The differentially expressed genes and its copy number variation between HO-8910PM cell line and normal ovarian tissues was detected by human genome UI33A 2. 0 gene chip and human mapping 10K array 2. 0 gene chip, and the data was analyzed by bioinformatics. Some of metastasis-associated genes were validated the results of single nucleotide polymorphism （SNP） and cDNA chips by fluorescence in situ hybridization （FISH） and real-time quantitative PCR. Results Integrate analysis of two gene chips data showed that there were 385 differentially expressed genes in the same and 379 SNP positional point （6 of them, included 2 genes） between HO-8910PM cell line and normal ovarian tissues, these copy number amplification of 379 SNP positional point of chromosome were ≥3, which had 240, deletion ≤ 1 had 139. Chromosome location analysis showed that there were 385 differentially expressed genes located at all chromosomes, and 261 of them （67.8%, 261/385） located at 10 chromosomes, included that 34 （8.8%）, 33 （8.6%）, 28（7.3%）, 27 （7.0%）, 25 （6.5%）, 24 （6.2%） of them located at chromosome 3, 2, 9, 10, 1 and 11 respectively, and 23 （6.0%） of them at chromosome 6 and 12 each, 22 （5.7%） of them at chromosome 4 and 5 each. For the function of differentially expressed genes, the results showed that 99 （25. 7% ） genes belonged to the family of enzymes and their regulators, 54 （ 14. 0% ） genes associated with signal transduction, 50 （ 13.0% ） genes associated with nucleic acid binding, and 36 （ 9. 4% ） genes associated with protein binding. Conclusion We have demonstrated that there are 4 kinds of differentially expressed genes related to metastasis of ovarian cancer, which belonged to the families enzyme and its regulator, nucleic acid binding, signal transduction and protein binding, and located at chromosome 1, 2, 3,4, 5, 6, 9, 10, 11 and 12.