Identification and characterization of alternative splicing sites of ADAR2 pre-mRNA in human glioma cell lines

• 1、Department of Neurosurgery, China–Japan Union Hospital, Jilin University
• 2、College of life science, Zhejiang Chinese Medicine University

Abstract：Objective To identify the alternatively splicing sites for ADAR2 pre-mRNA in glioma cell lines U87, U251, A172 and normal human astrocyte HA1800. Methods Premers were designed according to the alternatively splicing sites reported in reviews and divided the total cDNA of ADAR2 into six superposed fragments. ADAR2 mRNA was amplified by reverse transcription-polymerase chain reaction (RT-PCR). The PCR products were purified and then ligated into pGEM-T Easy vector. After transformation, positive clones were identified by PCR and sequencing. Results We identified three alternatively splicing sites in glioma cell lines U87, U251, A172 and normal human astrocyte HA1800: the first one located between exon -1 and 1 and created exon 1a; the second one results in the splicing-out of the whole of exon 2, which encodes both dsRNA-binding domains; and the third located in the deaminase domain and created exon5a. Conclusion There are three alternatively splicing sites of ADAR2 in glioma cell lines U87, U251, A172 and normal human astrocyte HA1800.

Keywords： Surgery Glioma ADAR2 Alternative splice Cloning