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目的:检测结肠癌细胞株中两种代谢相关基因DPYD和MTHFR的单核苷酸多态性(single nucleotide polymorphisms,SNPs)。方法:培养HT-29及Lovo两株结肠癌细胞,分别提取基因组DNA,PCR扩增DPYD和MTHFR基因相应目的片段,采用基于基质辅助激光解吸电离飞行时间质谱技术进行DPYD基因rs1801159(A/G)、rs1801160(G/A)、rs17376848(A/G),MTHFR基因rs1801131(A/C)、rs1801133(C/T)、rs2274976(G/A)的SNPs位点检测。结果:两种细胞的DPYD基因3个位点rs1801159、rs1801160及rs17376848基因型均为野生纯合型,即A/A、G/G及A/A型。HT-29细胞MTHFR基因rs1801131和rs1801133、rs2274976位点基因型分别为A/C、C/T杂合子和G/G纯合子,而Lovo细胞此3个位点依次为A/A、T/T纯合子和G/A杂合子。结论:本实验中HT-29及Lovo两种细胞株的DPYD基因所测3个SNP位点基因型均相同,而这两种结肠癌细胞株MTHFR基因所测的各SNP位点基因型均不相同。
Objective: To detect single nucleotide polymorphisms (SNPs) of DPYD and MTHFR in two colon cancer cell lines. Methods: The two colon cancer cell lines, HT-29 and Lovo, were cultured and genomic DNA was extracted respectively. The DPYD gene rs1801159 (A / G) was amplified by PCR-based amplification of DPYD and MTHFR gene fragments by matrix assisted laser desorption / ionization time- , Rs1801160 (G / A), rs17376848 (A / G), MTHFR rs1801131 (A / C), rs1801133 (C / T) and rs2274976 (G / A) Results: The genotypes of rs1801159, rs1801160 and rs17376848 in the three loci of DPYD gene were wild homozygous, ie A / A, G / G and A / A genotypes. The genotypes of rs1801131, rs1801133 and rs2274976 in MTHFR gene of HT-29 cells were A / C, C / T heterozygote and G / G homozygote respectively. The three sites of Lovo cells were A / A, T / T Homozygotes and G / A heterozygotes. Conclusion: The genotypes of three SNP loci detected by DPYD in both HT-29 and Lovo cell lines are the same in this experiment, but none of the genotypes of SNPs detected by MTHFR in these two colon cancer cell lines the same.