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Purpose: In this study, expression, genetic/epigenetic alterations, and biomarker potential of DNAJB11 gene were evaluated in breast cancer.Methods: DNAJB11 mRNA levels were analyzed in breast cell lines by qRT-PCR. In-silico tools (UALCAN and Kaplan-Meier plotter) were used to analyze the transcript level, prognostic survival value, and promoter methylation status of DNAJB11 in clinical breast cancer samples. Methylation status of DNAJB11 promoter region in cell lines was analyzed by Methylation Specific PCR (MSP).Results: DNAJB11 mRNA expression is significantly high in breast cancer cell lines compared to the non-tumorigenic breast cell line (MCF 10A). Clinical breast cancer samples (n = 1097) have significantly high levels of DNAJB11 transcripts compared to normal samples (n = 114) (P = 1.62e-12). High DNAJB11 mRNA levels were associated with poor overall survival (n = 626) (P = 0.042), relapse-free survival (n = 1764) (P = 1.3e-09) and distant metastasis free-survival (n = 664) (P = 0.00025) in breast cancer. Genetic alterations of DNAJB11 are infrequent in breast cancer samples. DNAJB11 promoter region was frequently hypomethylated in breast cell lines and breast cancer patient samples.Conclusion: It could be suggested that epigenetic mechanisms other than DNA methylation could be involved in the DNAJB11 regulation. DNAJB11 has unfavorable survival prognostic value in breast cancer.