摘要
目的:研究小核核糖核蛋白多肽N基因(SNRPN)在肝癌肿瘤HepG2细胞株的表达及基因印迹状态.方法:采用RT-PCR方法检测出SNRPN基因在肝癌肿瘤HepG2细胞株中的表达状况,对HepG2细胞株基因组DNA和cDNA中的SNRPN基因外显子4nt1654312位点用RT-PCR为基础的RFLP方法进行基因分型.结果:HepG2细胞稳定表达SNRPN,SNRPN外显子4nt1654312(数字依据NT_026446,SNPrs705)为杂合子(C/T);RT-PCR为基础的RFLP分析表明,SNRPN的双等位基因中只有T等位基因产生mRNA转录本.结论:SNRPN基因在HepG2肝癌细胞株中有表达,其基因印迹状态未丢失.
AIM: To investigate the expression and imprinting style of small nuclear ribonucleoprotein polypeptide N (SNRPN) in hepatic cancer cell line HepG2.
METHODS: Human hepatic cancer cell line HepG2 was cultured in vitro by routine method. The expression of SNRPN gene in the cells was detected by reverse transcription polymerase chain reaction (RT-PCR). The single nucleotide polymorphisms (SNP) of SNRPN at exon 4 nt 1654312 (numbered according to NT_026446, SNP rs705) was genotyped in a genomic DNA sample and a cDNA sample of HepG2 cell line with restriction fragment length polymorphism (RFLP) based on RT-PCR.
RESULTS: SNRPN was stably expressed in HepG2 cells. The heterozygote C/T was found at exon 4 nt 1654312 of SNRPN. In cell lines heterozygous with respect to this SNP, only one of the two alleles (T allele) present in the genomic DNA produced an mRNA transcript.
CONCLUSION: SNRPN mRNA is expressed in HepG2 cells, and there is no loss of imprinting.
出处
《世界华人消化杂志》
CAS
北大核心
2005年第21期2545-2548,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30470964~~
关键词
小核核糖核蛋白多肽N
基因印迹
限制性片段长度
多态性
杂合子
Small nuclear ribonucleoprotein polypeptide N
Genomic imprinting
Restriction fragment length polymorphism
Heterozygote