We determined the effects of organ environment on the response of murine CT‐26 colon carcinoma cells to 2 structurally and pharmacologically distinct chemotherapeutic agents. CT‐26 cells were injected i.v. (to produce lung lesions), s.c., into the cecal wall, and into the spleen (to produce spleen and liver lesions). Doxorubicin (DXR) at 10 mg/kg, 5‐fluorouracil (5‐FU) at 20 mg/kg, or saline (control) was injected intravenously on different schedules after tumor‐cell implantation. The in vivoresponses of the tumors growing in the cecum, spleen, liver, lung and subcutis were compared. Colon carcinomas growing in the subcutis were most sensitive to DXR. Tumors growing in the spleen and cecum were most sensitive to 5‐FU and less so to DXR. Tumors in the liver were highly resistant to both drugs, whereas experimental lung metastases were sensitive to 5‐FU but resistant to DXR. The differential responses of the tumors to the drugs were not due to drug distribution. The level of protein‐kinase‐C activity was elevated in the spleen, liver and cecum tumors as compared with s.c. tumors and correlated with the in vivo DXR resistance of the tumor cells. This correlation suggested that organ environment may modulate the chemosensitivity of tumor cells, at least in part, by perturbing signal transduction pathways. Collectively, the data indicate that the organ environment has profound effects on the response of tumor cells to chemotherapy. A molecular understanding of this phenomenon should facilitate the design of more effective systemic chemotherapy for cancer metastases. © 1992 Wiley‐Liss, Inc.