激光显微切割技术用于子宫内膜异位组织DNA的提取及其完整性分析

摘要

目的：：探索一套激光显微切割（LCM）分离子宫内膜异位症腺体细胞后提取微量DNA并进行完整性分析的操作流程。方法：分别对20例石蜡标本及20例冰冻标本进行LCM，收集切割后的腺体细胞；2组标本各取10例提取微量DNA，检测DNA浓度并通过PCR扩增进行验证；余20例标本分别进行全基因组扩增，检测产物浓度并利用8种常见管家基因作为引物通过PCR扩增进行验证，对比分析其结果。结果：石蜡标本与冰冻标本在LCM获取腺体细胞及提取微量DNA两个环节中均可获得满意效果；但经全基因组扩增后，石蜡标本无法保留完整DNA信息。结论：LCM获取子宫内膜异位症腺体细胞提取微量DNA是一种操作简单、结果稳定的方法，可作为日后子宫内膜异位症基因组研究的常规方法；冰冻切片相对石蜡切片，更能保留完整的DNA信息。%Objective: To detect a standard protocol for isolation of low amount DNA from the endometriotic cells obtained by laser capture microdissection(LCM). Methods: Using LCM to collect endometriotic cells from 20 paraffin specimens and 20 frozen specimens. 10 cases of each group were used to extract DNA. The others were used to do whole genome amplification. The production was validated by spectrophotometer and PCR was per-formed with housekeeping genes in different chromosomes. Results: Both the frozen samples and formalin-fixed par-affin embedded tissue can extract DNA production. However, after whole genome amplification, paraffin specimens can not remain intact DNA information. Conclusion: Isolation the genomic DNA from endometriotic cells obtained by LCM is a simple and stable method, which can be used as a routine approach for the future genome research of endometriosis.