Effect of TNF-α rs1800620 and rs4645843 on the Content of TNF-α in LX-2 Cell Line / 中国药师

ABSTRACT

Objective: To construct the cell models of TNF-α rs1800620 (SNP1) and TNF-α rs4645843 (SNP2) stable transgen-ic lines and study the effects of SNPs on the content of TNF-α in LX-2 cell line. Methods: TNF-α SNP1/TNF-α SNP2 mutant plasmid was synthesized by wild-type TNF-α targeting and then transfected into LX-2 cell line after lentivirus packaging. RT-qPCR and Western blot were used to detect the mRNA and protein levels of TNF-α in each recombinant cell. Results: The gene sequencing showed that plasmids were successfully constructed. Photographs from an inverted microscope showed that the recombinant cells had stable expres-sion of GFP. The results of RT-qPCR and Western blot showed that compared with the wild group, SNP1 and SNP2 could significantly reduce the mRNA (P<0. 05) and protein (P<0. 05) levels of TNF-α. Conclusion: The recombinant cells stably expressing TNF-α/TNF-α SNP1/TNF-α SNP2 LX-2 are constructed. SNP1 and SNP2 can significantly reduce the mRNA and protein levels of TNF-α in LX-2 cell line.