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Objective:To investigate the role of Caspase-1/gasdermin D (GSDMD) -mediated cell pyroptosis in anti-tumor effect of cisplatin (DDP) in triple-negative breast cancer (TNBC) .Methods:HE staining and immunohistochemical staining were performed to detect the morphological changes and the expression of pyroptosis/apoptosis pathway related proteins in TNBC tissues before and after DDP-based neoadjuvant chemotherapy (NACT) . The TNBC cell line MDA-MB-231 was treated with DDP and the morphological changes were observed. The type of cell death induced by DDP was analyzed by Annexin V-FITC/PI double staining and flow cytometry. Lactate dehydrogenase (LDH) release assay and ELISA were performed to detect the release of LDH and inflammatory factors (IL-18 and IL-1β) in cell culture supernatant after DDP treatment. Western blot (WB) was performed to detect the expression of pyroptosis/apoptosis pathway related proteins in cells after DDP treatment. MDA-MB-231 cells treated with DDP were co-treated with caspase-1 specific inhibitor to inhibit pyroptois or co-treated with caspase-3 specific inhibitor to inhibit apoptosis. The effect of caspase-1 inhibitor or caspase-3 inhibitor on the anti-tumor effect of DDP was detected by MTT assay, clone formation assay, transwell assay and would healing test.Results:Reactive changes in the breast surgical specimen after DDP-based NACT included cell swelling and inflammatory cell aggregation around the tumor bed, which were more similar to pyroptosis. The up-regulation of key molecules of pyroptosis pathway post-NACT was significantly higher than that of key molecules of apoptosis pathway. Further experiments in vitro showed that DDP could induce MDA-MB-231 cells to show pyroptosis-like changes characterized by large bubbles blowing from the cellular membrane. Flow-cytometry analyses showed that the death type of MDA-MB-231 cells caused by DDP was mainly Annexin V +PI + cells (mainly lytic cells, such as pyroptosis) . Additionally, DDP treatment induced significant activation of caspase-1 and GSDMD, increased the release of LDH, IL-18 and IL-1β, however, the activation level of caspase-3, which dominates the apoptosis pathway, was significantly lower than that of caspase-1/GSDMD. Moreover, caspase-1 inhibitors (blocking the classical pyroptosis pathway) had a significantly greater inhibitory effect on the anti-tumor effect of DDP than caspase-3 inhibitors (blocking the apoptosis pathway) . Conclusion:Caspase-1/GSDMD mediated pyroptosis may play a leading role in the anti-tumor effect of DDP in triple-negative breast cancer.
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Objective To discuss the clinical pathological features,diagnosis and differential diagnosis,therapy and prognosis of primary breast diffuse large B-cell lymphoma(PB-DLBCL).Methods The clinical manifestations,pathological features,immunophenotypic characteristics of 4 cases of PB-DLBCL were retrospectively reviewed,and related literatures were reviewed.Results All of the 4 patients were women,aged 45 to 69 years,with the median 58.5 years.There were 3 cases of lesions involving the left breast and 1 case of the right breast.Microscopic observation showed that the normal structure of the mammary gland was destroyed,and no capsule was seen.There was a large number of large-to medium-sized heterotypic lymphoid cells which characterized as diffuse infiltration between the remaining ducts or lobules of mammary gland.Tumor cells infiltrated adipose tissue,mainly as central blast-like cells (3 cases) and immunoblast-like cells (1 case).No lymphatic epithelial lesions were observed.The immunophenotype showed 4 cases were non-GCB type.Immunostaining showed that the neoplastic cells were LCA,CD20,CD79a and MUM-1 positive.Ki67 index were from 50％ to 80％.The follow-up time of these cases was until Dec.2017.There was one recurrence during the follow-up period.The patient died 13 months later.One patient was alive and had survived for 66 months.Two patients were lost to follow-up.Conclusions PB-DLBCL is extremely rare,mainly found in women.Most of the cell origin types are non-GCB type,which is often misdiagnosed as breast cancer.The diagnosis of PB-DLBCL is confirmed by pathological biopsy and immunohistochemical markers.The treatment is not clear,and a comprehensive treatment plan such as R-CHOP/ CHOP chemotherapy combined with radiotherapy is recommended.