The aim of this study is to investigate the expression of sarcosine metabolism related proteins according to androgen receptor (AR) and HER-2 status in estrogen receptor (ER) negative breast cancer and to analyze its clinical implications. Tissue microarray was constructed for a total of 334 cases of ER negative breast cancer. Immunohistochemical stain was conducted for sarcosine metabolism related proteins such as glycine N-methyltransferase (GNMT), sarcosine dehydrogenase (SARDH), and l-pipecolic acid oxidase (PIPOX). There were 131 AR positive, 205 AR negative cases and 143 HER-2 positive, 193 HER-2 negative cases. When subdividing into four groups according to AR and HER-2 status, there were 55 AR(+)/HER-2(-) cases, 76 AR(+)/HER-2(+) cases, 67 AR(-)/HER-2(+) cases and 138 AR(-)/HER-2(-) cases. GNMT and PIPOX expression was highest in the AR(+)/HER-2(-) group while expressed lowest in the AR(-)/HER-2(-) group (P<0.001). Stromal PIPOX expression was highest in the AR(-)/HER-2(+) group and lowest in the AR(-)/HER-2(-) group (P=0.010). GNMT and PIPOX expression was higher in the AR positive group compared with those of AR negative group (P=0.001, and P<0.001, respectively), while tumoral and stromal PIPOX expression showed a significant association with HER-2 positivity (P=0.006, and P=0.005, respectively). AR positive group had the highest ratio of low sarcosine type while the AR negative group had the highest ratio of null type (P<0.001). In conclusion, ER negative breast cancer showed different expression of sarcosine metabolism related proteins according to AR and HER-2 status. GNMT and PIPOX expression was high in the AR positive group while tumoral and stromal PIPOX expression was high in the HER-2 positive group.