Chemotherapy contained anthracycline inevitably cause declines of cardiac function. This study evaluated the deterioration of left ventricular segmental systolic function in patients with lymphoma received anthracycline chemotherapy detected by echocardiographic three dimensional speckle tracking imaging (3D-STI). Sixty patients with newly diagnosed diffuse large B-cell lymphoma who received R-CHOP chemotherapy were enrolled. Three dimensional left ventricular global longitudinal strain (3D-GLS), three dimensional left ventricular global circumferential strain (3D-GCS) and three dimensional left ventricular longitudinal strain of different left ventricular segments (3D-LS) were measured by 3D-STI at baseline, after the completion of two cycles and four cycles of the regimen respectively. Compared with baseline, 3D-GLS reduced significantly after four cycles of anthracycline therapy (P < 0.001), while 3D-GCS showed no significant variation during the whole procedure (all P > 0.05). For individual segment, LS of apical anterior and septal walls decreased significantly after two cycles of chemotherapy (all P < 0.05). After four cycles of treatment, 3D-LS of the mid-ventricular level (all P < 0.05), apical level (all P < 0.05) and apex (P < 0.05) worsened. Serum hs-cTnT levels increased after anthracycline exposure (P < 0.05) and Serum hs-cTnT levels were correlated with 3D-GLS at the end of four cycles (r = 0.12, P = 0.03). Mean values of involved segmental 3D-LS of two and four cycles were both correlated with serum hs-cTnT levels at the end of both two and four cycles (r = 0.368, P = 0.041; r = 0.79, p < 0.001). 3D-STI evaluation of the LV provides an understanding of the segmental impairment of LV wall and the possible process of LV impairment in lymphoma patients after anthracycline chemotherapy.