Objective: Microtubule plays a critical role in oncogenesis and progression of breast cancer, which is mediated by tubulin gene superfamily to a great extent. The current research aims to systematically analyze expression levels and prognostic values of tubulin gene superfamily members in breast cancer and identify novel functional microtubule genes. Methods: The Cancer Genome Atlas (TCGA) and the Kaplan-Meier plotter database were used to define the expression levels and prognostic values of tubulin gene superfamily members in breast cancer, respectively. Expression pattern and prognostic value of TUBA1C was subsequently confirmed on breast cancer tissue microarray (TMA) by immunochemistry (IHC) staining. Moreover, cell viability and apoptosis assays were conducted in MDA-MB-231 and MCF-7 breast cancer cells to investigate the functional role of TUBA1C in breast cancer. Results: Systematic analysis of tubulin gene superfamily revealed that TUBA1C was significantly overexpressed and had notable prognostic value in breast cancer. IHC analysis exhibited that the TUBA1C was overexpressed at the protein level in tumor tissues compared with para-tumor tissues (χ2=6.929, P=0.008) and upregulated TUBA1C was associated with the advanced clinical stage (χ2=6.357, P=0.042) and worse overall survival (OS) (P=0.021). In addition, univariate (P=0.032) and multivariate COX regression analyses (P=0.040) further demonstrated that TUBA1C was an independent prognostic factor in breast cancer. Besides, Knockdown of TUBA1C notably suppressed cell proliferation and induced cell apoptosis. Conclusion: These results revealed that TUBA1C is a novel oncogene and a potential prognostic biomarker in breast cancer. Besides, silencing TUBA1C could inhibit breast cancer progression, which could be a potential target for therapy.