Smad4 protein, whose gene is coded at chromosome 18q21.1, is an important tumour suppressor that mediates transforming growth factor-beta. It has been reported that inactivation of the Smad4 gene and allelic loss of chromosome 18q correlate with liver metastasis and poorer prognosis in colorectal cancers. Utilising a recently developed method of immunohistochemical staining for Smad4 protein, we focused on the specific impact of Smad4 protein expression on liver metastasis in colorectal cancer. We also evaluated the association between chromosome18q deletion and liver metastasis. We selected 20 colorectal cancers with liver metastasis for the experimental group, and 20 cases without liver metastasis for the control. In order to exclude the influence of lymph node metastasis, all cases were lymph node negative. In addition, the two groups were matched for tumour depth, tumour differentiation and tumour location. We compared the expression level of Smad4 protein immunohistochemically in these 20 matched pairs. We also compared the loss of heterozygosity status at chromosome 18q in these 20 matched pairs. Immunohistochemical staining revealed a significant difference (P ¼ 0.024) in the level of Smad4 protein between the two groups. We also observed a significantly different (P ¼ 0.0054) ratio of allelic deletion at chromosome 18q21. Smad4 protein expression level and allelic loss at 18q21 are associated with the process of liver metastasis in colorectal cancers evaluated when excluding clinical and pathological features except for liver metastasis. (Vogelstein et al, 1988(Vogelstein et al, , 1989Benhattar et al, 1993;Kinzler and Vogelstein, 1996;Span et al, 1996;Kambara et al, 2004;Nassif et al, 2004). Loss of heterozygosity (LOH) is one of the major types of genetic inactivation, and the long arm of chromosome 18 is the most frequently deleted region in colorectal cancers. To date, many reports suggest that this deletion is a molecular predictor that affects survival (Fearon et al, 1990;Jen et al, 1994;Chung, 1998;Lanza et al, 1998;Ogunbiyi et al, 1998;Jernvall et al, 1999;McLeod and Murray, 1999;Sarli et al, 2004). We too reported that the allelic deletion of chromosome 18q was associated with poorer prognosis in stage III colon cancer after adjuvant chemotherapy (Watanabe et al, 2001. These reports suggest that there might be tumour suppressor genes located at chromosome 18q, which has a strong influence on survival. Smad4 gene, which mediates the intracellular signalling pathway of transforming growth factor (TGF)-beta receptor, has been detected as one of the target genes at 18q21 (Wang et al, 1995;Eppert et al, 1996;Hahn et al, 1996;Thiagalingam et al, 1996;Carethers et al, 1998;Markowitz, 2000;Fink et al, 2003;Alazzouzi et al, 2005;Li et al, 2005). Recently, an immunohistochemical method for evaluating Smad4 protein has been developed and several studies found higher frequency of Smad4 protein inactivation in the cases with liver metastasis and in the cases presenting unfavourable survival (Maitra et al, 20...