In this study we examined traditional prognostic factors and a panel of protein markers associated with breast cancer progression, aggressiveness, hypoxic response and cell invasion/metastasis, respectively NHERF1, VEGFR1, HIF-1α and TWIST1, to determine whether they are differentially expressed in tumors scored as grade 2, trying to improve their prognosis definition. The gene discussed is TWIST1; the disease is breast cancer.