The classification of germinal centre (GC) and non-GC is an important prognostic immunophenotype for patients with diffuse large B cell lymphoma (DLBCL) following anthracycline-based chemotherapy. The expression of the anti-apoptotic protein, Bcl-2, has been associated with an unfavourable prognosis in patients with DLBCL. Immunohistochemistry was performed using antibodies against CD10, Bcl-6, MUM-1 and Bcl-2. To establish the combined prognosis of the immunophenotype and Bcl-2 overexpression, patients were separated into four subgroups based on their gene expression profile: the Bcl-2 positive GC subgroup, Bcl-2 negative GC subgroup, Bcl-2 positive non-GC subgroup, and the Bcl-2 negative non-GC subgroup. The clinical characteristics and survival outcomes of the four patient subgroups were compared. Ninety-six patients with de novo DLBCL received R-CHOP (rituximab, cyclophosphamide, vincristine, adriamycin and prednisone) therapy. The baseline characteristics of the patient subgroups were similar. The Bcl-2 negative GC subgroup had a favourable progression-free and overall survival (OS) compared with the other three subgroups (p = 0.042, 0.043). Multivariate analysis confirmed that the Bcl-2 negative GC group was independently associated with progression-free survival and OS. The results of this study showed that the Bcl-2 negative GC patients had the most favourable prognosis among patients with DLBCL that received R-CHOP.