Blocking a single gene interferes with the metabolism of vitamin D and cancer trails in the new cell examination

Last study published in the journal He examined the role of an enzyme with short -chain dehydrogenase/reductase 42E (SDR42E1) in regulation of sterol metabolism and vitamin D.

Vitamin D is a Pleotropic hormone necessary for calcium phosphorus and homeostasis, bone immunity and health function. Deficiencies may result from impaired absorption and metabolism, despite the exposure to sunlight and the availability of dietary sources. The receptor of vitamin D (VDR) mediates in the biological effect of vitamin D, while the enzyme from the cytochrome family P450 24 (CYP24A1) The enzyme regulates its inactivation.

Association testing throughout the vitamin D deficiency gene showed a nonsense variant RS11542462, which produces a non -functional SDR42E1 enzyme. This variant correlates with an increased level of 7-dehydrocholesterol (7-DHC) and 8-DHC, precursors in vitamin D synthesis. In addition, the authors previously identified these sterols as potential SDR42E1 substrates; However, the exact role of the enzyme in the metabolism of vitamin D remains unknown.

Examination and results

In this study, scientists have explained the role of SDR42E1 in regulating the metabolism of sterol and vitamin D. First, they introduced plasmid containing SDR42E1 into HCT116 cells, a cellular lines of human colorectal cancer (CRC), in order to analyze its expression and cell distribution. Western Blot analysis confirmed the expression of SDR42E1 with the expected molecular weight. The assessment of cellular location was revealed by its presence on the cell membrane and cytoplasm.

Then, the shortened short palindrome repetitions/protein 9 related to CRISPR (CRISPR/CAS9) were used regularly to generate a homozygous Knock-in model with RS11542462 to generate a homozygic model Knock-in mutation To assess the functional role of SDR42E1 in HCT116 cells. RNA sequencing analysis revealed 4663 genes of various expression (DEG) in the Knock-in model compared to wild type cells.

About 56% DEG was adjusted up with a short (FC) ≥ 0.3 change, including low -density (LDLR) 1B (LRP1B), protein casset (ABC), ATP cassette (ABC). In addition, 525 degrees were downwards; Some of them are aldolaza fructose-bisfosphate A (Aldoa), 2H3A histone cluster (Hist2H3A), a member of the WNT 16 (WNT16) family and a member of the dissolved family 7 (SLC7A5).

The encyclopedia of genes and genomes in Kyoto (KEGG) identified many activated routes, including digestion and absorption of vitamin, inflammatory intestinal disease, immune disorders, biosynthesis of cholelish acid of primary gile acid and ABC transporters. However, routes related to cellular aging, replication and DNA repair, regulation of the cell cycle and improper regulation of transcription in cancer have been deactivated.

In addition, the specific changes of genes were recorded on the routes of cellular aging and cancer, including a reduction in the heat shock of 90 Alpha Family Family Member 1 (HSP90AA1) and WNT2 and regulation up to human leukocytes up antigen A (HLA-A) as well as a signal transducer and transcription activator 2 (Stat2). In addition, LRP1 and ABCA1 were adjusted up, and the 24-DHC (DHCR24), CYP51A1 and LDLR reductase were adjusted down in the routes related to steroid biosynthesis and cholesterol metabolism.

Then, the analysis of liquid chromatography was revealed by 140 proteins of various expression in the Knock-in model compared to wild type cells. Adjustable proteins included ribosomal protein L11 (RPL11), Hist2H3A, Histone 1 H4A (Hist1H4A) and a family member H2A Y Histone (H2AFY). Down-adjustable proteins included kinase 12 (APAP12) anchoring protein, Synthetase Acyl-coa long chain member 5 (ACSL5), syntase of fatty acids (FASN), Aldoa and Aldoc.

In addition, the SDR42E1 was hyperextered in HCT116 cells to examine its impact on sterol trails and biological profiles. This revealed significant transcription deregulation, with 1483 degrees compared to controls. Of these, 343 genes were adjusted up and 95 were adjusted down. Paths related to the immune response, cellular aging, cholesterol metabolism, improper regulation of transcription in cancer and signaling of the Kappa B nuclear factor were regulated up in the SDR42E1 overexpressions model.

On the other hand, anti -phalan resistance, DNA replication and repair, and collecting conduit acid release routes were adjustable down in the hypertension model. In addition, significant genetic changes were observed in trails related to cholesterol metabolism, steroid biosynthesis, cancer and cellular aging. The authors also note that SDR42E1 overexpression induced transcriptional changes related to the reaction to stress and apoptotic resistance, including the Ad regulation up the PRO-SURVIVAL and pro-metastatic genes, such as HSPA5, CXCL8 and Serpine1. Finally, scientists examined the role of SDR42E1 in cell life. They found a significant reduction in cell life in HCT116 SDR42E1 cells, which showed that the study can be reversed through the transitional overall overall.

Conclusions

To sum up, the discoveries emphasize the SDR42E1 as a key modulator in trails related to vitamin D. The disturbance of SDR42E1 affects the absorption regulators of vitamin D, metabolic homeostasis and cell proliferation. It also affects several characteristic features, especially those related to CRC progression and chemotherapy reaction. And vice versa, SDR42E1 overexpression caused a change in transcription in accordance with apoptotic resistance and reaction to stress. The article also suggests that the SDR42E1 may play a wider role in modulating vitamin D signaling both genomic and non -emergency (initiated by the membrane), justifying further research.

Study restrictions include the use of HCT116 cells that may not replicate the functional and structural complexity of vitamin D, the potential of effects outside the goal with the CRISPR/CAS9 gene and lack of complementary functional tests. In addition, the cancerous origin and limited potential for differentiation of HCT116 cells affect the generalization of the results. Further research is necessary to confirm these results and better therapeutic and physiological assessment of SDR42E1.