应用抑制性消减杂交筛选卵巢癌紫杉醇耐药差异表达基因

目的:应用抑制性消减杂交技术研究人卵巢癌紫杉醇耐药细胞株OC3/Tax300与其亲本细胞OC3(敏感株)之间基因表达的差异,筛选耐药相关基因,探讨基因表达差异与卵巢癌耐药的相关性。方法:以卵巢癌紫杉醇耐药细胞株OC3/Tax300 mRNA为检测子(tester),以其亲本细胞OC3(敏感株)mRNA为驱赶子(driver),构建cDNA消减文库。随机挑取文库克隆测序,所得结果在GenBank中做同源性比较分析。结果:成功构建了卵巢癌紫杉醇耐药细胞株的特异性cDNA消减文库。从文库中选取阳性克隆,其中76个测序成功,再随机选取36个序列与GenBank数据库进行初步比对,8个可能为新基因,1个为蛋白序列,另27个来源于16个已知基因,这些差异表达基因主要涉及细胞代谢、信号转导、细胞骨架、凋亡、蛋白翻译合成、发育等相关基因。结论:部分基因表达差异与卵巢癌紫杉醇耐药有关。

Screening differentially expressed genes of Taxol-resistant in ovarian cancer by suppres-sion subtractive hybridization

Objective:To investigate the differential gene expression of taxol-resistant in ovarian cancer cell line OC3/Tax300 compared with its parental cell OC3(sensitive lines) using suppression subtractive hybridization;to study its relationship with Drug-resistance and its mechanism.Method:To construct subtractive cDNA library of taxol-resistant ovarian cancer cell line from the mRNA of OC3/Tax300(tester) and OC3(driver),partial positive clones in the library were randomly selected,sequenced and then analyzed with the BLAST software in GenBank.Result:Subtractive cDNA library of taxol-resistant ovarian cancer cell line was constructed with 76 clones successfully sequenced,among which 36 clones were selected randomly and compared with the GenBank Database.27/36 sequence had a high similarity to 16 known genes;1/36 was protein sequence,while 8/36 failed to match with the homologous genes in Genbank.These subtractive genes were mainly related to cellular energy process,signaling pathway,cytoskeleton,apoptosis,protein synthesis,growth and development,etc.Conclusion:Part of these differentially expressed genes is related to the mechanisms of taxol-resistance of ovarian cancer.