论文部分内容阅读
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 dihydropyrimidine dehydrogenase (DPYD) and methylenetetrahydrofolate reductase (MTHFR) gene in gastric cancer cell lines. Methods The genomic DNAs of BGC-823, SGC-7901, MKN-28, AGS and MCG were extracted by QIAamp DNA Blood Mini kit. The primers were designed and the corresponding fragments were amplified by PCR. Matrix-assisted laser desorption / ionization time of flight mass spectrometry Technology for DPYD gene rs1801159, rs1801160, rs17376848 and MTHFR gene rs1801131r, s1801133r, s2274976 SNPs loci detection. Results The genotype of rs1801159 in DPYD gene of AGS cell line was G / A, and the rest of the cell lines were A / A homozygous. The genotypes of rs1801160 and rs17376848 in DPYD gene of the five cell lines were G / G and A / A. AGS cell line MTHFR gene rs1801131 site A / C heterozygotes, the rest of the cell lines are A / A homozygous AGS cell line MTHFR gene rs1801133 locus genotype C / C, the remaining cell lines of this site All of which were C / T heterozygotes; the rs2274976 sites of MTHFR gene of five cell lines were all G / G homozygotes. Conclusion The rs1801159 locus of DPYD gene and the rs1801131 and rs1801133 sites of MTHFR gene of AGS cell line are different from the other four cell lines. The other loci of the five gastric cancer cell lines have the same genotype.