HepG2和Huh7细胞CYP3A4、CYP3A5和PXR基因非编码单核苷酸多态性分析

Analysis of noncoding single nucleotide polymorphism of CYP3A4,CYP3A5 and PXR gene in HepG2 and Huh7 cells

目的:对HepG2和Huh7细胞中CYP3A4、CYP3A5和PXR基因的重要非编码单核苷酸多态性(SNP)进行分析。方法:提取HepG2和Huh7细胞基因组DNA,PCR后进行测序分析。结果:在HepG2细胞中CYP3A4 rs2242480、rs3735451、rs2246709、rs4646440、rs12333983都为突变杂合子;而在Huh7细胞中上述前4个SNP为野生纯合子,最后1个为突变纯合子。在这两种细胞中,CYP3A4 rs35599367、rs4646437、rs35564277都为野生纯合子,rs2740574为突变纯合子。在HepG2和Huh7细胞中CYP3A5 rs776746分别为突变杂合子和突变纯合子。在两种细胞中,PXR rs3814055、rs2276706、rs6785049为突变杂合子,rs13059232为突变纯合子。上述14个SNP中,HepG2细胞突变杂合子较多,而Huh7细胞野生纯合子及突变纯合子较多。结论:HepG2和Huh7细胞中CYP3A4、CYP3A5 SNP基因型不完全相同,而PXR SNP基因型相同。

Aim:To analyze important noncoding single nucleotide polymorphism(SNP)of CYP3A4,CYP3A5 and PXR gene in HepG2 and Huh7 cells.Methods:Cellular genomic DNA in HepG2 and Huh7 cells was extracted,and sequencing analysis was performed after PCR.Results:In HepG2 cell,CYP3A4 rs2242480,rs3735451,rs2246709,rs4646440,and rs12333983 were all mutant heterozygotes,while the first 4 SNPs were wild homozygotes,and the last SNP was mutant homozygote in Huh7 cell.CYP3A4 rs35599367,rs4646437 and rs35564277 were wild homozygotes,while rs2740574 was mutant homozygote in these two cells.CYP3A5 rs776746 in HepG2 and Huh7 cells were mutant heterozygote and mutant homozygote,respectively.PXR rs3814055,rs2276706,rs6785049 were mutant heterozygotes,and rs13059232 was mutant homozygote.Among the above 14 SNPs,HepG2 cells had more mutant heterozygotes,while Huh7 cells had more wild homozygotes and mutant homozygotes.Conclusion:The genotypes of CYP3A4 and CYP3A5 SNPs in HepG2 and Huh7 cells are not exactly the same,while the genotypes of PXR SNPs are the same.