中德临床肿瘤学杂志（英文版）2005，Vol.4Issue(5)：309-313,5.
激光捕获显微切割结合RNA线性扩增技术在膀胱移行细胞癌相关基因研究中的应用
Laser Capture Microdissection Combined with RNA in vitro Linear Amplification Detecting the Relevant Genes of Bladder Cancer
郝权 1徐勇 1李文录 1赵小鸽 1刘文欣1
作者信息
- 1. 天津医科大学附属肿瘤医院盆腔科 300060
- 折叠
摘要
Abstract
Objective: Laser capture microdisection has become indispensable to the analysis of the difference of gene expression between human bladder transitional cell and bladder transitional cell carcinoma (BTCC). However, to obtain sufficient RNA from laser-capture microdissected cells is quite difficult. The study was designed to determine a feasible technical routine to isolate transitional cells from bladder membrane, separate carcinoma cells from stromal cells and to amplify the RNA isolated from laser-capture microdissected cells. Methods: Bladder transitional cell were obtained from frozen sections of bladder membrane applying LCM, by the same token, BTCC cells from frozen sections of BTCC tissue. Then RNA was extracted and linearly amplified in vitro. The expression levels ofβ-actin in primary total RNA and amplified RNA were detected using RT-PCR. Results: That RNA integrity was good after LCM was confirmed by control experiment Ⅰ; By control experiment Ⅱ, the correlation between the number of LCMshooting and RNA quantity under arranged conditions was preliminarily confirmed. About 0.5-2.5kb RNA fragments were obtained after RNA amplification and β-actin levels were integral. Conclusion: Laser capture microdissection combined with RNA linear amplification in vitro can be successfully applied to obtain pure objective cells for research. The integrity of the amplified RNA is good and can be employed in further research.关键词
激光捕获显微切割/RNA体外线性扩增/RT-PCR/膀胱移行上皮细胞/膀胱癌细胞Key words
aser capture microdissection/RNA linear amplification in vitro/RT-PCR/bladder transitional cell/bladder transitional cell carcinoma分类
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郝权,徐勇,李文录,赵小鸽,刘文欣..激光捕获显微切割结合RNA线性扩增技术在膀胱移行细胞癌相关基因研究中的应用[J].中德临床肿瘤学杂志（英文版）,2005,4(5):309-313,5.
