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目的高通量地筛选人卵巢癌相关的自然反义转录子,为深入研究自然反义转录子在卵巢癌发生、发展中的作用奠定基础。方法以人正常卵巢组织和卵巢癌组织为材料,用RNase I保护分析法分离出两种组织中的正-反义转录子双链RNA,经逆转录后用抑制性消减杂交特异性扩增两种组织间的差异片段,最后对差异片段克隆、测序。结果分离出的两种组织的正-反义转录子经克隆和鉴定后,共获得20个测序结果,其中有3个分子与卵巢癌等肿瘤可能密切相关,说明该方法可用于肿瘤相关NAT分子的系统筛选,并为深入研究卵巢癌相关NAT分子提供了靶标。
Objective To screen natural antisense transcripts related to human ovarian cancer with high throughput and to lay a foundation for the further study on the function of natural antisense transcripts in the occurrence and development of ovarian cancer. Methods Human normal ovarian tissue and ovarian cancer tissue were used as materials. The double-stranded RNA of sense-antisense transcripts in both tissues was isolated by RNase I protection assay. After reverse transcription, The difference between the species of the fragment, the final fragment of the difference cloning, sequencing. Results The positive and negative antisense transcripts of the two tissues were cloned and identified. Totally 20 sequencing results were obtained, of which 3 were closely related to the tumors such as ovarian cancer, indicating that this method can be applied to tumor-associated NAT molecules System screening, and provide a target for further study of ovarian cancer-associated NAT molecules.