Neuroblastoma breakpoint family member 1 (NBPF1) is involved in the occurrence and development of tumors. However, only a limited number of studies were conducted on NBPF1 and cutaneous squamous cell carcinoma (SCC). This study mainly explored the expression and mechanism of NBPF1 in SCC. SCC tissue and adjacent tissues samples were randomly selected. NBPF1 gene was overexpressed in the A431 cell line using plasmid transfection technique. Cell viability was tested by cell counting kit-8 (CCK-8) assay. Flow cytometry was used to determine cell cycle and apoptosis. Western blot and RT-qPCR were respectively performed to determine the expression levels of proteins and mRNAs. The NBPF1 gene was lowly expressed in SCC tissues. The expression level of NBPF1 gene was the lowest in A431 cell line. The cell viability of A431 was reduced after transfection. Overexpression of NBPF1 not only arrested A431 cells in G1 phase and promoted apoptosis, but also up-regulated the expressions of Bax and p53 mRNA and protein and down-regulated the expressions of Bcl-2, Survivin and Cyclin D1. Akt-p53-Cyclin pathway was inhibited when NBPF1 gene expression was up-regulated. Upregulation of NBPF1 might promote apoptosis of A431 cells and block cell cycle via inhibiting the activation of Akt-p53-Cyclin signaling pathway.