Abstract Expression of the proliferating cell nuclear antigen (PCNA) was examined in normal rat intestinal tissues and in carcinogen-treated nonneoplastic and neoplastic colonic mucosa. In the normal intestine, PCNA expression was confined to the expected region of the proliferative compartment. A strong correlation was observed between PCNA labeling index and both [ 3H]thymidine labeling index ( R = 0.993, P = 0.007) and percent of cells in S phase as determined by flow cytometry ( R = 0.982, P = 0.018) and between the location of the maximal staining for PCNA and [ 3H]thymidine ( R = 0.997, P < 0.05). In animals treated with dimethylhydrazine (DMH), crypt hyperplasia, an increased PCNA labeling index, and shifts in both the region of maximal and the upper extent of PCNA expression were observed during DMH exposure; significant crypt hyperplasia and expansion of the PCNA-positive compartment persisted after completion of DMH injections. The patterns of PCNA expression and bromodeoxyuridine incorporation were similar in DMH-induced tumors. It is concluded that PCNA immunohistochemistry can be used as a reliable marker of the proliferative compartment in both normal and neoplastic colonic mucosa.