Cancer is a complex disease that involves the alterations of metabolic pathways and tumor microenvironment. Lactate in the tumor microenvironment leads to cancer proliferation, metastasis, and angiogenesis. However, the effect of lactate on prostate cancer cells is still unclear. Here, our objective is to identify the significant molecules and biological processes by analyzing the RNA-seq data. The GSE195639 was produced by the Illumina NextSeq 500 (Homo sapiens). The KEGG and GO analyses show that Herpes simplex virus 1 infection and Rap1 signaling pathway are considered major pathways during the lactate-treated cancer cells. Furthermore, we identified the top ten essential molecules including IL6, CASP3, JUN, MAPK3, BRCA1, PIK3R1, CCNA2, TPI1, APOE, and EXO1. Therefore, our study may provide novel insights into the mechanism of prostate cancers.