[Splice variants of mRNA of cytochrome P450 genes: analysis by the nanopore sequencing method in human liver tissue and HepG2 cell line]
- PMID: 35485485
- DOI: 10.18097/PBMC20226802117
[Splice variants of mRNA of cytochrome P450 genes: analysis by the nanopore sequencing method in human liver tissue and HepG2 cell line]
Abstract
The analysis of cytochrome P450 transcripts was carried out by the nanopore sequencing in liver tissue samples of three donors and HepG2 line cells. It has been demonstrated that direct mRNA sequencing with a MinION nanopore sequencer (Oxford Nanopore Technologies) allows one to obtained quantitative profiles for transcripts (and their splice variants) of cytochrome P450 superfamily genes encoding isoforms involved in metabolism of the large (~80%) part of drugs. The splice variant profiles substantially differ for donors. The cytochrome P450 gene expression at the transcript level is significantly weaker in cells of the HepG2 line compared with that in the normal liver tissue. This limits the capability of the direct mRNA nanopore sequencing for studying alternative splicing of cytochrome P450 transcripts in HepG2 cells. Both quantitative and qualitative profiles of the cytochrome P450 gene expression at the transcript level are notably differ in human liver tissue and HepG2 cells.
Metodom nanoporovogo sekvenirovaniia proveden analiz transkriptov tsitokhromov R450 v obraztsakh tkani pecheni trekh donorov i kletkakh linii HepG2 (gepatoblastoma cheloveka). Pokazano, chto priamoe sekvenirovanie mRNK s ispol'zovaniem nanoporovogo sekvenatora MinION (Oxford Nanopore Technologies) pozvoliaet poluchat' kolichestvennye profili transkriptov (i ikh splaĭs-variantov) dlia genov supersemeĭstva tsitokhroma R450, kodiruiushchikh izoformy, pri uchastii kotorykh metaboliziruetsia bol'shaia chast' (~80%) lekarstvennykh veshchestv. Profili splaĭs-variantov sushchestvenno razlichaiutsia dlia raznykh donorov. Ékspressiia genov tsitokhromov R450 na urovne transkriptov v kletkakh linii HepG2 znachitel'no slabee v sravnenii s ikh ékspressieĭ v normal'noĭ tkani pecheni, chto zatrudniaet issledovanie al'ternativnogo splaĭsinga transkriptov tsitokhromov R450 v kletkakh HepG2 metodom priamogo nanoporovogo sekvenirovaniia mRNK. Profili ékspressii genov supersemeĭstva tsitokhroma R450 na urovne trankriptov v tkani pecheni cheloveka i kletkakh HepG2 znachitel'no razlichaiutsia kak kolichestvenno, tak i kachestvenno.
Keywords: HepG2 cell line; cytochromes P450; human liver tissue; nanopore sequencing; splice variants.