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背景与目的:当对无间质细胞混杂的肿瘤细胞进行选择性分析时,显微切割技术是不可缺少的,然而,从显微切割所得的细胞中获取足够RNA量相当困难。本文的目的是寻找一特异方法,将鼻咽癌细胞从肿瘤间质细胞中分离出来,并对其扩增,以备进一步研究。方法:采用显微切割技术从鼻咽癌组织冰冻切片中获取鼻咽癌细胞,提取RNA并对微量RNA进行体外转录,然后,用RT-PCR分别验证扩增RNA的β-actin和GADPH基因表达水平。结果:从鼻咽癌组织中获取了20000～40000个细胞,抽提RNA,扩增后获得了片断大小为0.5～2.5kb的RNA,在β-actin、GADPH表达完整。结论:显微切割结合RNA线性扩增技术能成功地获取较为单一的鼻咽癌细胞,扩增后的RNA完整性较好,能运用于进一步研究中。
BACKGROUND & AIM: Microdissection is essential for the selective analysis of mesenchymal-free tumor cells. However, obtaining sufficient amounts of RNA from microdissected cells is quite difficult. The purpose of this paper is to find a specific method to separate nasopharyngeal carcinoma cells from tumor stromal cells and amplify them for further study. Methods: Nasopharyngeal carcinoma cells were obtained from frozen sections of nasopharyngeal carcinoma by microdissection. RNA was extracted and transcribed into microRNAs in vitro. Then, the expression of β-actin and GADPH gene in amplified RNA was verified by RT-PCR Level. RESULTS: From 20000 to 40000 cells were obtained from NPC tissues and RNA was extracted. The RNA of 0.5-2.5kb in size was obtained after amplification. The expression of β-actin and GADPH was complete. Conclusion: Microdissection combined with RNA linear amplification technology can successfully obtain more single nasopharyngeal carcinoma cells. The amplified RNA has better integrity and can be used for further research.