Aim - To explore the role and possible mechanism of KNTC1 gene in the development of bladder cancer. Methods - The expression level of KNTC1 in bladder cancer tissues and adjacent tissues was detected by immuno-histochemistry. Mann-Whitney U, Spearman, and Kaplan-Meier analyses were used to analyze the correlation between KNTC1 expression level and various characteristics of bladder cancer cases. KNTC1 was knocked down by RNA interference to detect the proliferation, apoptosis, cell cycle, migration, and in vivo tumorigenesis of bladder cancer cells. Human apoptosis antibody array and Western blot were used to detect the expression level changes of related proteins after KNTC1 knockdown to explore the possible mechanism. Results - KNTC1 was highly expressed in bladder cancer tissues and was related to the pathological grade and overall survival rate of bladder cancer. Knockdown of KNTC1 can inhibit the proliferation, migration, and in vivo tumorigenesis of bladder cancer cells and promote apoptosis. KNTC1 knockdown changes the levels of many proteins in bladder cancer cells, including Caspase 3, Fas, p-Akt, CDK6, PIK3CA, and MAPK9. Conclusion - KNTC1 plays a crucial role in the development of bladder cancer, and our findings provide evidence for its use as a therapeutic target.