CYP2W1 promotes experimental metastatic-colonization phenotypes and is associated with cholesterol-biosynthesis-related molecular alterations in colorectal cancer

BACKGROUNDColorectal cancer (CRC) commonly spreads to the liver, and this metastatic disease accounts for much of the mortality associated with CRC.Its development involves several linked changes, among which are epithelial-mesenchymal transition and metabolic adaptation.Cholesterol metabolism is one such change.The oncofetal monooxygenase cytochrome P450 family 2 subfamily W member 1 (CYP2W1) is frequently detected in CRC and has been related to poor outcome, but whether it has a functional role in liver metastasis is not known. AIMTo examine the contribution of CYP2W1 to CRC experimental liver-colonization phenotypes and cholesterol-biosynthesis-related molecular alterations. METHODSExpression data from public databases were considered together with findings from 18 paired clinical specimens.Stable knockdown cells were prepared from CT26, HCT116, and SW480, and a CT26 overexpression model was also established.Cell growth was examined by cell counting kit-8 and colony formation; wound-healing and Transwell assays were used for motility.Subcutaneous implantation, intrasplenic injection, and tailvein injection provided three animal models (n = 6 mice per experimental group).Reverse transcription quantitative real-time polymerase chain reaction and immunoblotting were used to examine cholesterol synthesis molecules and protein kinase B (AKT)-mechanistic target of rapamycin (mTOR) signaling, while low-density lipoprotein receptor was assessed as an indicator of cholesterol uptake. RESULTS / 44Liver metastases contained more CYP2W1 than primary CRC tissues.High CYP2W1 expression was associated with shorter survival in Kaplan-Meier Plotter (hazard ratio = 1.36,P = 0.0037) and The Cancer Genome Atlas datasets (hazard ratio = 2.98, P = 0.00163).Removing CYP2W1 reduced CRC cell growth, colony formation and migration.The same manipulation slowed subcutaneous tumors and decreased liver and lung experimental metastatic-colonization in mice (n = 6 mice per group); overexpression produced the reverse pattern.Knockdown cells had less intracellular cholesterol, along with lower HMGCS1, HMGCR, nuclear SREBF2, phosphorylated AKT, and phosphorylated mTOR.Low-density lipoprotein receptor did not change consistently.Cholesterol supplementation restored part of the lost proliferative and migratory related phenotypes.In the 18 paired tissue cohort, CYP2W1, HMGCS1, and SREBF2 staining was stronger in liver metastases than in primary tumors. CONCLUSIONOur pre-clinical data demonstrate that CYP2W1 modulates CRC proliferative and experimental metastatic-colonization phenotypes.These changes are associated with altered AKT-mTOR-SREBF2 signaling and cholesterol-biosynthesis-related molecular markers, no definitive causal ordering was established.This correlative metabolic relationship warrants further mechanistic validation before therapeutic translation for metastatic CRC.