【摘要】 目的:摸索一条可行的技术路线,运用激光捕获显微切割技术(lasercapturemicrodissection,LCM)获取无间质混杂的人正常子宫内膜腺管上皮细胞与子宫内膜癌细胞,用于进行子宫内膜癌相关基因差异表达的研究。方法:采用LCM技术分别从正常子宫内膜组织及子宫内膜癌组织冰冻切片中捕获子宫内膜腺管上皮细胞及子宫内膜癌细胞,提取微量RNA,并对微量RNA进行纯化及浓缩,用RTPCR验证捕获细胞RNA中β-actin基因表达水平。结果:分别从冰冻切片中捕获子宫内膜腺管上皮细胞及子宫内膜癌细胞各50000Shootings。对照实验Ⅰ、Ⅱ证实经LCM后RNA完整性较好。确定了LCMShooting次数与可获得RNA量间对应关系。捕获细胞所提取的RNA中βactin基因表达完整。结论:通过LCM技术可以从冰冻切片中捕获单一类型的目的细胞,此过程不会造成细胞RNA明显降解,可以应用于下游的实验研究。更多还原

【Abstract】 OBJECTIVE:To find a feasible technique route to acquire pure human normal endometrial glandular epithelial cells and endometrial carcinoma cells by laser capture microdisection (LCM) and apply it in the research of endometrial carcinoma relevant gene’s diffrerent expressions. METHODS:Pure endometrial glandular epithelial cells and endometrial carcinoma cells from tissue frozen sections were obtained by applying LCM, then minimal RNA was extracted, purified and concentrated. The expression level of β-actin in RNA of LCM cells was detected by using RT-PCR. RESULTS:Each 50 000 shottings of endometrial glandular epithelial cells and endometrial carcinoma cells were captured by LCM. The RNA integrity was well confirmed by control experimentⅠand Ⅱ. The correlation between the LCM Shootings and RNA quantity under arranged conditions was preliminary conformed. β-actin expression was integral in RNA extracted from LCM cells. CONCLUSION:Laser capture microdissection can be apllied to successfully obtain pure objective cells from frozen sections. The integrity of RNA is good after LCM process and can be used in downstream experiment.更多还原