CENPF overexpression facilitates the G1/S cell cycle transition of hepatocellular carcinoma cells via MYC pathway

Centromere protein F (CENPF), a mitosis-related protein, is overexpressed in hepatocellular carcinoma (HCC) and has emerged as a promising biomarker for early HCC. However, with ultra-large molecular weight of 358kDa of CENPF, no study has directly explored its carcinogenicity with an overexpression model. In the present study, a CRISPR/dCas9 system was applied to construct the CENPF overexpression model. CENPF was upregulated and downregulated to analyze its functions in vitro. CENPF knockdown cell models showed inhibition of HCC proliferation. Notably, as a cell cycle protein with high constitutive expression in G2/M phase, CENPF overexpression cell models also showed inhibitory effects, probably due to the toxic effect of excessive CENPF expression on G2/M transition. However, in both CENPF downregulation and overexpression models, cell cycle assays showed CENPF promoted G1/S transition in HCC cells. RNA-seq showed that CENPF overexpression activated the MYC pathway, thereby promoting G1/S transition. The rescue experiment indicated that the MYC pathway inhibitor 10058-F4 counteracted the G1/S transition induced by CENPF overexpression in HCCLM3 cells. CENPF overexpression might facilitate HCC cells G1/S cell cycle transition via the MYC pathway.