Objective To study the expression of CD24, G protein coupled receptor 5 (Lgr5) and Ki-67 in colorectal cancer tissues and their clinical importance, in order to provide guidance for clinical diagnosis and treatment of colorectal cancer. Methods The clinical datas of 200 cases patients with colorectal cancer admitted to The First People's Hospital of Fangchenggang from January 2018 to December 2020 were analyzed retrospectively, and 200 specimens of colorectal cancer tissues and 200 specimens of mucosa tissues adjacent to of colorectal cancer in the same case were collected. All specimens were detected by immunohistochemistry to compare the positive expression rates of CD24, Lgr5 and Ki-67 in colorectal cancer tissues and mucosa tissues adjacent to of colorectal cancer , and the relationship between the expression of CD24, Lgr5 and Ki-67 and the clinicopathological characteristics of colorectal cancer patients were analyzed. Results The positive expression rates of CD24, Lgr5 and Ki-67 in colorectal cancer tissues were significantly higher than those in mucosa tissues adjacent to colorectal cancer; among the patients with CD24 positive expression in colorectal cancer tissues, the proportion of patients with≥40 years old, liver metastasis and low differentiation of colorectal cancer were significantly higher than those patients with CD24 negative expression; among the patients with Lgr5 positive expression in colorectal cancer tissues, the proportion of patients with low differentiation was significantly higher than that patients with Lgr5 negative expression; among the patients with Ki-67 positive expression in colorectal cancer tissues, the proportion of colorectal cancer patients with liver metastasis and low differentiation were significantly higher than those patients with Ki-67 negative expression (all P <0.05). Conclusion CD24, Lgr5 and Ki-67 are highly expressed in colorectal cancer tissues, and the expression levels of CD24, Lgr5 and Ki-67 are closely related to the clinicopathological characteristics of patients, which can be used as the important index in the diagnosis and treatment of colorectal cancer.