Fresh frozen breast cancer tissue from every third patient diagnosed and treated between 1991 and 2004 at the Koo Foundation Sun-Yat-Sen Cancer Center (KFSYSCC) were randomly selected for the study. Patients with follow-up periods shorter than three years were excluded, with the exception of those who died of the disease within three years of the initial treatment. In cases of ineligibility, the following sample was selected. The selected tissue samples spanned the major transition periods of adjuvant chemotherapy from CMF (cyclophosphamide, methotrexate and fluorouracil) to CAF (cyclophosphamide, doxorubicin, fluorouracil) and to taxane-based regimens. Four hundred forty seven samples were obtained, but 135 samples were excluded due to insufficient RNA (n = 1), poor RNA quality (n = 116), or unacceptable microarray quality (n = 18). A total of 312 samples were eligible for the study (Cohort 1). Gene expression profiles of an additional 15 lobular breast carcinoma samples, collected between 1999 and 2004 and previously studied, were also included (Cohort 2). All patients were treated by a multidisciplinary team according to the guidelines consistent with the National Comprehensive Cancer Network [18 ]. Following modified radical mastectomy or breast-conserving surgery plus dissection of axillary nodes, patients received radiotherapy, adjuvant chemotherapy, and/or hormonal therapy, if indicated. Neoadjuvant chemotherapy was administered to patients with locally advanced disease. The study was approved by the institutional review board (ID number 20020128A) and ethical approval was obtained from the same board for samples without obtainable informed consent.
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