One‐quarter of a century ago, identification of the human retinoblastoma gene (RB) loci proved Knudson's ‘two‐hit theory’ that tumor suppressor genes exist. Since then, numerous works delineated crucial roles for the RB protein (pRB)‐E2F transcription factor complex in G1‐S phase transition. In addition, discovering the relationship between pRB and tissue‐specific transcription factors enabled a better understanding of how cell cycle exit and terminal differentiation are coupled. Recent works provoked many exciting twists in views on pRB functions during cancer initiation and progression beyond its previously well‐appreciated roles. Various mitogenic and cytostatic cellular signals appeared to modulate pRB functions and thus affect a wide variety of effector molecules. In addition, genetic studies in mice as well as other creatures incessantly force us to revise our views on pRB functions. This review will focus particularly on the roles of pRB in regulating intracellular signaling, cell metabolism, chromatin function, stem cells and cancer stem cells. (Cancer Sci 2012; 103: 1182–1188)