[Application of microdissection combined with RNA linear amplification].
OBJECTIVE
Microdissection has become indispensable for the selective analysis of stroma-free tumor cell. However, To obtain sufficient RNA from microdissected cells is difficult. The study was designed to seek a specific way to separate nasopharyngeal carcinoma (NPC) cells from stromal cells and to amplify the RNA from microdissected NPC cells.


METHODS
NPC cells were obtained using microdissection from frozen NPC tissue sections, then RNA was extracted from the microdissected NPC cells and reverse transcribed in vitro. The expression levels of beta-actin and GADPH in amplified RNA were detected using RT-PCR.


RESULTS
About 20,000-40,000 NPC cells were obtained, RNA was extracted from the cells, about 0.5-2.5 kb RNA fragments were obtained after RNA linear amplification and beta-actin and GADPH levels were integral.


CONCLUSIONS
Microdissection combined with RNA linear amplification can be used to successfully obtain pure NPC cells, the integrity of amplified RNA is good and can be used in further research.
Han
Wei-nong
WN
Key Laboratory of Chinese Ministry of Education, Xiangya School of Medicine, Central South University, Changsha 410078, P. R. China.
Li
Hong
H
Wang
Shuang
S
Zhou
Wen
W
Yao
Kai-tai
KT
D004210:Dissection / Q000379:methods; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000235:genetics* / Q000473:pathology; D012334:RNA, Neoplasm / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012334:RNA, Neoplasm