摘要
目的检测胃癌细胞株中二氢嘧啶脱氢酶(DPYD)、亚甲基四氢叶酸还原酶(MTHFR)基因的单核苷酸多态性(SNPs)。方法采用QIAamp DNA Blood Mini kit提取BGC-823、SGC-7901、MKN-28、AGS及MCG五种胃癌细胞株基因组DNA,设计引物,PCR扩增相应目的片段,采用基质辅助激光解吸电离飞行时间质谱技术进行DPYD基因rs1801159、rs1801160、rs17376848及MTHFR基因rs1801131r、s1801133r、s2274976的SNPs位点检测。结果 AGS细胞株DPYD基因rs1801159位点基因型为G/A,其余细胞株该位点均为A/A纯合子;五种细胞株DPYD基因rs1801160及rs17376848位点基因型分别为G/G及A/A。AGS细胞株MTHFR基因rs1801131位点为A/C杂合子,其余细胞株该位点均为A/A纯合子;AGS细胞株MTHFR基因rs1801133位点基因型为C/C,其余细胞株该位点均为C/T杂合子;五种细胞株MTHFR基因rs2274976位点均为G/G纯合子。结论 AGS细胞株DPYD基因rs1801159位点及MTHFR基因rs1801131、rs1801133位点与其余四种细胞株不同,五种胃癌细胞株其它所测位点基因型相同。
Objective To detect single nucleotide polymorphisms（SNPs） of dihydropymidine dehydrogenase（DPYD） and methylenetetrahydrofolate reductase（MTHFR） genes in gastric cancer cell lines.Methods The genomic DNA of BGC-823,SGC-7901,MKN-28,AGS and MCG cell lines was isolated by QIAamp DNA Blood Mini kit.The primes were designed and the related target DNA fragments were amplified by PCR.The SNPs genotyping of DPYD gene rs1801159,rs1801160,rs17376848 and MTHFR gene rs1801131,rs1801133,rs2274976 was performed by means of matrix-assisted laser desorption ionization time-of-flight mass spectrometry method（MALDI-TOF-MS）.Results The genotype of DPYD gene locus rs1801159 was G/A in AGS cell line,which was A/A in the other 4 cell lines.The genotypes of DPYD gene loci rs1801160 and rs17376848 were G/G and A/A,respectively,in the 5 detected cell lines.The genotype of MTHFR gene locus rs1801131 was A/C in AGS cell line,which was A/A in the other 4 cell lines.The genotype of MTHFR gene locus rs1801133 was C/C in AGS cell line,which was C/T in the other 4 cell lines.The genotype of MTHFR gene locus rs2274976 was G/G in the 5 detected cell lines.Conclusion The locus rs1801159 of DPYD gene and loci rs1801131,rs1801133 of MTHFR gene are different in AGS cell line from the other 4 cell lines,while the other detected loci of the 2 genes are similar in the 5 detected gastric cancer cell lines.
出处
《江苏医药》
CAS
CSCD
北大核心
2011年第17期2000-2003,I0001,共5页
Jiangsu Medical Journal
关键词
胃癌
单核苷酸多态性
二氢嘧啶脱氢酶基因
亚甲基四氢叶酸还原酶基因
Gastric cancer
Single nucleotide polymorphisms
Dihydropymidine dehydro-genase gene
Methylenetetrahydrofolate reductase gene
