Background: We made investigations on the expression of negative costimulatory molecules B7-H1, B7-H3, B7-H4 in lesions at various stages of human colorectal cancer (CRC) evolution. We also analyzed the relevance between its expression in CRC and pathological factors in clinic and patient survival time.We finally made a result that we can ensure the clinical significance of B7-H3, B7-H4 expression and the relationship between the B7 family molecules expression.Methods: All stages of CRC were collected, including polyps, adenomas, high-grade neoplasms, and colorectal carcinomas. There were 98 cases of resected CRC tissue, 30 cases of polyps, 30 cases of adenomas, and 25 cases of high-grade neoplasia. Then analysis the expression of three negative costimulatory molecules in all stages of colorectal tissue expression patterns, flow cytometry CD3 + T lymphocytes B7 family molecules to explore the potential value of their expression.Results: The expression of B7 family molecules in 30 cases of polyps, 30 cases of adenomas, 25 cases of highgrade neoplasia, and 98 cases of CRC tissues revealed that the B7-H1 and B7-H3 expression quantity was found in polyps, adenomas, high-grade neoplasms, as well as they were highly expressed in cancerous tissues. B7-H4 was only expressed in cancerous tissues. In the TNM stage of tumor progression, three negative costimulatory molecules were mainly expressed in the cytoplasm of tumor cells. In CRC in lymphocytes B7-H4 expression was related to patient age, mucinous adenocarcinoma, and lymph node metastasis. Survival analysis showed in CRC it was statistically significant that that the expression of B7-H3 and the survival rate of patients. We found that 92.9% of CRC patients expressed B7 family negative costimulatory molecules in varying degrees, and the prognosis and co-expression of B7 family negative co-stimulatory molecules were negatively correlated.Conclusions: Costimulatory molecules B7-H1, B7-H3 were expressed on the early stage of CRC development. The amount of CD3 + T lymphocyte infiltration in CRC was positively correlated with the survival of patients. They are the earliest molecules to participate in the progress of CRC developing in B7 family negative molecules. This suggests that B7-H1 and B7-H3 are involved in rectal carcinoma at the junction, as well as it is essential throughout the entire evolutionary process. However, it is in CRC tissues that B7-H4 only get expressed, and different expression patterns may have different clinical significances.