A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells

Abstract

Background: Cytochrome P450 2S1 (CYP2S1) is an orphan P450 with an unknown biological function. Data from our laboratory and others suggest that CYP2S1 may have an important physiological role in modulating the synthesis and metabolism of bioactive lipids including prostaglandins and retinoids. CYP2S1 expression is elevated in multiple epithelial-derived cancers as well as in the chronic hyperproliferative disease psoriasis. Whether CYP2S1 expression in proliferative disease is protective, detrimental, or neutral to disease progression remains to be determined. Two human bronchial epithelial cells (BEAS-2B) were constructed to represent chronic depletion of CYP2S1 using short-hairpin RNA (shRNA) silencing directed toward the 3'UTR (759) and exon 3 (984) of the CYP2S1 gene and compared with a non-targeting shRNA control (SCRAM). Both CYP2S1 mRNA and protein were depleted by approximately 75% in stable cell lines derived from both targeted shRNA constructs (759 and 984). To elucidate the biological significance of CYP2S1, we analyzed transcriptome alterations in response to CYP2S1 depletion in human lung cells. Results: RNA-sequencing (RNA-seq) analysis was performed to compare the transcriptome of the control (SCRAM) and the CYP2S1-depleted (759) BEAS-2B cell lines. Transcriptomes of the replicates from the two cell lines were found to be distinct populations as determined using Principal Component Analysis and hierarchical clustering. Approximately 1000 genes were differentially expressed in response to CYP2S1 depletion. Consistent with our previous phenotypes, DAVID analysis revealed altered regulation in key pathways implicated in cell proliferation and migration. Transcriptomic profiles were also consistent with the metabolism of proposed endogenous substrates. Pathway analysis also revealed significant expression changes within mTOR signaling, a critical pathway in cell growth. To determine whether these changes manifest as altered cell size, cell diameter and volume were calculated, revealing that CYP2S1 depletion promotes cell growth in BEAS-2B cells. Conclusions: These data suggest that pathway analysis of sequence-based gene expression is a powerful method to identify pathways and phenotypic alterations in response to changes in orphan enzyme expression. Our results suggest a novel role for CYP2S1-mediated metabolism in modulating BEAS-2B cell size. These findings warrant further studies on CYP2S1 regulated pathways to elucidate potential substrates of CYP2S1.

Development of a KRAS-Associated Metabolic Risk Model for Prognostic Prediction in Pancreatic Cancer

“…Indeed, CYP2S1 affected the metabolism of arachidonic acid (AA) and linoleic acid (LA) to modulate BEAS-2B cells growth. [17] In consistent with their results, our data also found CYP2S1 was enriched in Lipid metabolic process. It will be interesting to investigate the detailed mechanism by which CYP2S1 regulate lipid metabolism in PC in future study.…”

Development of a KRAS-Associated Metabolic Risk Model for Prognostic Prediction in Pancreatic Cancer

“…Study showed that expressions of 8 CYPs were changed in CYP2S1-depleted cells, and all of them were involved in lipid biotransformation. Indeed, CYP2S1 affected the metabolism of arachidonic acid (AA) and linoleic acid (LA) to modulate BEAS-2B cell growth [ 18 ]. In consistent with their results, our data also found CYP2S1 was enriched in lipid metabolic process.…”

CYP2S1 rs338599 polymorphism confers reduced risk to anti‐tuberculosis drug‐induced liver injury and may be a novel marker for its risk prediction

“…CYP2S1 mRNA expression level is low in liver, but high in extrahepatic tissues such as respiratory and digestive systems. 10 In addition, CYP2S1 mRNA and protein are also expressed in human skin. CYP2S1 plays an important physiological role in the metabolism of dodecanoids, and may play a role in inflammatory response, tumor, and other pathological processes.…”

“…13.2, including 9 exons, encoding 504 amino acid proteins with molecular weight of about 55.8 kDa, with 8 single‐nucleotide polymorphisms (SNPs) loci. CYP2S1 mRNA expression level is low in liver, but high in extrahepatic tissues such as respiratory and digestive systems 10 . In addition, CYP2S1 mRNA and protein are also expressed in human skin.…”