卵巢癌相关基因的筛选与鉴定

背景与目的:卵巢癌在妇科肿瘤中死亡率最高,但是目前对于在卵巢癌发生中的敏感和特异癌基因或抑癌基因的改变还知之甚少。因此,筛选新的卵巢癌相关基因并对其作用进行研究可能有助于探讨卵巢癌发生的机理,并可为卵巢癌的早期诊治提供有意义的生物学指标。本研究旨在获得新的卵巢癌相关基因。方法:在改良mRNA差异显示方法的基础上,结合反向Northern点杂交等鉴定方法,筛选卵巢癌组织和对侧正常卵巢组织之间的差异表达基因,进一步利用生物信息学资源对所获得的差异表达基因片段进行分析。并以差异表达片段作为探针,原位杂交方法检测基因在其它卵巢正常组织和卵巢癌组织中的表达。结果:筛选后得到12个差异表达基因,其中3个为基因组重复序列,5个未知基因序列,4个已知基因序列。原位杂交结果表明锌指蛋白361(Zincfingerprotein361,ZNF361)基因、蛋白酶体PSMA2(proteasomesubunit,alphatype2,PSMA2)基因、位于人1号染色体的未知基因OCRC13和位于9号染色体的未知基因OCRC4在36例卵巢癌组织中的表达均高于其在16例正常卵巢组织中的表达(P<0.05)。尤其是与鼠配子形成期特异表达家族成员-纺锤体蛋白(spindlin)基因高度同源的OCRC4基因,在16例正常卵巢癌组织中未见其表达,而在卵巢癌组织中的表达可达53%(17/36)。结

Screening and identification of ovarian carcinomas related genes

BACKGROUND &OBJECTIVE: Ovarian cancer is the leading cause of death among gynecological malignancies. Up to now, little is known about specific tumor suppressor genes or oncogenes involved in the ovarian cancer genesis. Thus isolation of new candidate genes and characterization of their role in ovarian cancer genesis will be helpful for understanding the molecular mechanisms and developing protocols for early diagnosis and therapy of ovarian carcinomas. This study was developed to screen and identify genes related to ovarian carcinomas. METHODS: Modified mRNA differential display PCR and reverse Northern dot blot analysis were used to screen and identify different displayed genes between ovarian carcinoma tissue and normal ovarian tissue. The genes were sequenced and analyzed by bioinformatics software. With these gene fragments as probes, in situ hybridization was used to characterize specific gene expression difference between ovarian cancer tissues and normal ovarian tissues. RESULTS: Of 12 differentially expressed genes, 5 were novel genes, 3 were chromosome genomic repeat sequences and the other 4 were known human genes. Furthermore, in situ hybridization analysis indicated that four genes, ZNF361, PSMA2, OCRC13 (a novel gene on chromosome 1) and OCRC4 (a novel gene on chromosome 9) were highly expressed in 36 samples of ovarian cancer tissues but not in 16 samples of normal ovarian tissues (P< 0.05). More interestingly, the gene OCRC4, with 99%sequence homology to mouse spindlin, a member of gene family specially expressed during gametogenesis, was highly expressed in 19 samples of ovarian cancer from total 36 cases (53%), but not expressed in control samples. CONCLUSION: Our observations indicate that the four genes ZNF361, PSMA2, OCRC4, and OCRC13 may be ovarian cancer related genes. It is speculated that novel gene OCRC4 might also be a member of specific gene family, which plays roles during early period of embryo development and tumorigenesis.