The androgen receptor (AR) has been demonstrated to have an impact on the effectiveness of metformin treatment, and typically, drug resistance develops in only a subset of cancer cells, not all cells.20,22 To explore whether a subgroup of DU145 cells with AR-negative expression is predisposed to acquiring metformin resistance, we performed single-cell RNA-seq analysis on a DU145 cell model undergoing the acquisition of metformin resistance (DU145 pre-MetR) and identified 6 clusters (0 through 5) using unsupervised clustering (Fig. 3l). This evidence concerns the gene AR and cancer.