Artesunate synergistically promotes sorafenib­induced apoptosis and ferroptosis in non­Hodgkin lymphoma cells through inhibition of the STAT3 pathway.

Despite the development of advanced therapies, the prognosis of non­Hodgkin lymphoma (NHL) remains unsatisfactory due to refractory and relapsed cases. Artesunate (ART) and sorafenib (SOR) both exert potential antitumor activity in lymphoma. The present study aimed to investigate whether ART and SOR produce synergistic anti­lymphoma effects, and to determine the potential underlying mechanisms. Cell viability assay, flow cytometry, malondialdehyde assay, GSH assay and western blotting were performed to evaluate cell viability, and changes in apoptosis, autophagic vacuoles, reactive oxygen species, mitochondrial membrane potential, lipid peroxidation and protein expression. The results demonstrated that ART and SOR synergistically inhibited the viability of NHL cells. ART and SOR also synergistically induced apoptosis, and markedly increased the expression levels of cleaved caspase­3 and poly (ADP­ribose) polymerase. Mechanistically, ART and SOR synergistically induced autophagy, and rapamycin enhanced the ART­ or SOR­induced inhibition of cell viability. Furthermore, it was demonstrated that ferroptosis promoted ART­ and SOR­induced cell death through increasing lipid peroxides. Erastin enhanced the inhibitory effects of ART and SOR on cell viability, whereas ferrostatin­1 reduced the ART­ and SOR­induced apoptosis of SU­DHL4 cells. Further studies revealed that signal transducer and activator of transcription 3 (STAT3) contributed to ferroptosis induced by ART and SOR in NHL cells, and genetic inhibition of STAT3 promoted ART/SOR­induced ferroptosis and apoptosis, concomitantly reducing the expression levels of glutathione peroxidase 4 and myeloid cell leukemia­1. Moreover, the combined treatment of ART and SOR exerted inhibitory effects on tumor growth, as well as antiangiogenic activity, resulting in the inhibition of CD31 expression in a xenograft model. Collectively, these findings indicated that ART acted synergistically with SOR to inhibit cell viability, and to induce apoptosis and ferroptosis through regulating the STAT3 pathway in NHL. Notably, ART and SOR may act as potential therapeutic agents for the treatment of lymphoma.

Treatment of refractory MALT lymphoma by lenalidomide plus bendamustine: A case report.

RATIONALE: Marginal zone B cell lymphoma of the mucosa-associated lymphoid tissue (MALT lymphoma) has an indolent natural course and disseminates slowly. However, there is currently no consensus regarding the optimal treatment strategy for relapsed/refractory MALT lymphomas. Lenalidomide-bendamustine may be an effective regimen for such cases. PATIENT CONCERNS: A 48-year-old Chinese male patient with MALT lymphoma and API2/MALT received 2 courses of standard-dose rituximab, cyclophosphamide, vincristine, prednisone regimen chemotherapy combined with Helicobacter pylori eradication therapy. However, this disease was not effectively managed. DIAGNOSIS: MALT lymphoma. INTERVENTIONS: The patient received lenalidomide-bendamustine (lenalidomide 25 mg on days 1-21 and bendamustine 90 mg/m2 on days 1-2) for 6 courses. OUTCOMES: Lenalidomide-bendamustine was a safe and effective chemotherapy. No serious adverse events occurred during the treatment period. Ultrasound gastroscopy revealed that the tumor gradually shrank and eventually disappeared to complete remission. LESSONS: The lenalidomide-bendamustine scheme might be a potentially effective option for patients with refractory or relapsed MALT lymphoma.

Artesunate induces apoptosis, autophagy and ferroptosis in diffuse large B cell lymphoma cells by impairing STAT3 signaling.

Artesunate (ART), a water-soluble derivative of artemisinin, has been reported to exert antineoplastic effects via diverse mechanisms in various types of cancer. Therefore, understanding the underlying mechanism of action of ART in distinct cancer types is indispensable to optimizing the therapeutic application of ART for different types of cancer. The present study aimed to investigate the cellular and molecular mechanisms responsible for the antineoplastic effects of ART in diffuse large B cell lymphoma (DLBCL) cells. Cell proliferation was measured using Cell Counting Kit-8 and colony formation assays. The levels of apoptosis and cell cycle distribution were investigated using flow cytometry. In addition, western blotting was used to analyze the expression levels of ART-induced apoptosis-, autophagy- and ferroptosis-related proteins. Monodansylcadaverine staining was performed to determine the levels of autophagy. Moreover, malondialdehyde and reactive oxygen species assays were used to determine the levels of ferroptosis. The results of the present study revealed that ART inhibited proliferation, and induced apoptosis, cell cycle arrest, autophagy and ferroptosis in DLBCL cells. Pharmacological inhibition of autophagy and ferroptosis alleviated the increased levels of apoptosis induced by ART. Notably, ART was found to exert its effects via inhibition of STAT3 activation. The genetic knockdown of STAT3 enhanced ART-induced autophagy and ferroptosis, and concomitantly upregulated the expression levels of apoptosis- and cell cycle-related proteins. In conclusion, the findings of the current study suggested that ART may induce apoptosis and cell cycle arrest to inhibit cell proliferation, and regulate autophagy and ferroptosis via impairing the STAT3 signaling pathway in DLBCL cells.

Causes of death and effect of non-cancer-specific death on rates of overall survival in adult classic Hodgkin lymphoma: a populated-based competing risk analysis.

BACKGROUND: The improved prognosis of classic Hodgkin lymphoma (cHL) has been accompanied by elevated risks of non-cancer-specific death (non-CSD). The aim of this study was to verify the occurrence of non-CSD and its effect on rates of overall survival among adult patients with cHL. METHODS: To ensure sufficient follow-up time, we analyzed retrospective data from patients aged ≥20 years with cHL that was diagnosed between 1983 and 2005 in the Surveillance, Epidemiology, and End Results (SEER) database. Logistic regression was applied to analyze the non-CSD occurrence in relation to all factors. Using Fine-Gray's method, we calculated the cumulative incidences of CSD and non-CSD. Stacked cumulative incidence plots and ratio of non-CSD to all causes of death were applied to evaluate the effect of non-CSD on rates of overall survival. Finally, we analyzed long-term mortality through Cox proportional hazard regression analysis and competing risk regression analysis to emphasize a more appropriate model of survival for patients with cHL. RESULTS: Among the 18,518 patients included, there were 3768 cases of CSD (20.3%) and 3217 of non-CSD (17.4%). Older age, earlier period, male sex, unmarried status, mixed cellularity (MC) and lymphocyte-depletion (LD) histological subtype, and patients received radiotherapy (RT) only were associated with more non-CSD according to binary logistic analysis. The cumulative incidence of non-CSD exceeded CSD after approximately 280 months follow-up. The most common causes of non-CSDs were cardiovascular disease, subsequent primary neoplasms, infectious diseases, accidents, and suicide. In a Cox proportional hazards model, patients who were black, unmarried, at an advanced stage or underwent chemotherapy (CT) alone were at greater risk of mortality than were white patients, who were married, at an early stage, and underwent combined modality; these populations were also found to be at greater risk for CSD in a competing risk model, but the risk of non-CSD did not differ significantly according to race and marital status, patients with early-stage disease and who underwent RT only were found to be at higher risk of non-CSD instead. CONCLUSIONS: Lymphoma was the cause of death in most patients who died, but non-CSD was not unusual. Patients with cHL should be monitored closely for signs of cardiovascular disease and malignant tumors. Rates of overall survival of patients were diminished by non-CSD, and a competing risk model was more suitable for establishing the prognosis than was the Cox proportional hazards model.

Treatment of refractory diffuse large B-cell lymphoma by chidamide combined with autologous stem cell transplantation: a case report.

Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma, with certain DLBCLs affecting specific anatomic sites, such as primary cutaneous DLBCL, leg type and intravascular large B-cell lymphoma. However, the occurrence of secondary cutaneous involvement in DLBCL while patients are undergoing regular chemotherapy is rare. In this study, we reported a case of refractory diffuse large B-cell lymphoma with cutaneous involvement that achieved complete remission for more than 4 years with epigenetic regulation of chidamide in combination with chemotherapy and autologous hematopoietic stem cell transplantation including a pretreatment regimen containing chidamide.

Aberrant TRPM4 expression in MLL-rearranged acute myeloid leukemia and its blockade induces cell cycle arrest via AKT/GLI1/Cyclin D1 pathway.

Transient Receptor Potential Melastatin Subfamily Member 4 (TRPM4) has been demonstrated to be aberrantly expressed in several cancers but seldom reported in acute leukemia. Based on database mining and validated experiments, our present data show that TRPM4 is selectively overexpressed in AML patients and cell lines with the MLL gene rearrangement. We analyzed the correlation between TRPM4 expression and clinical parameters in a validated cohort of AML patients. Increased TRPM4 expression was associated with significant leukocytosis (p = .028), M4/M5 subtype (p = .000), FLT3-ITD mutation (p = .034), MLL status (p = .007) and a higher risk stratification (p = .001). Knockdown of TRPM4 mediated by siRNA impaired proliferation and arrested the cell cycle at the G0/G1 phase in MLL-rearranged leukemia cells. We suggested that TRPM4 may be involved in the pathogenesis of MLL-rearranged leukemia through regulating the AKT/GLI1/Cyclin D1 pathway. The transcription factor HOXA9 was found to be responsible for upregulation of TRPM4 expression by binding to its promoter. In conclusion, TRPM4 is overexpressed in MLL-rearranged AML and blockade of TRPM4 may be an alternative therapeutic approach in AML patients with high TRPM4 expression.

[Expression and Significance of Pim-3 Gene in Acute Myeloid Leukemia].

OBJECTIVE: To investigate the role of Pim-3 abnomal expression in development of acute myeloid leukemia. METHODS: Semi-quantitative RT-PCR was used to detect the expression of Pim-3 in bone marrow of 47 newly diagnosed and untreated patients with acute myeloid leukemia (AML) and 18 patients with AML after treatment with chemotherapy. At the same time, the bone marrow of 10 cases of non-hematologic malignancies was used as normal control. The difference of the Pim-3 gene expression in bone marrows among the 3 groups was also compared. RESULTS: According to the RT-PCR detection results, the Pim-3 expression level in bone marrow of AML patients before chemotherapy were all significantly lower than those in patients with non-hematologic malignancies (P < 0.01). After chemotherapy, there were no significant differences of the Pim-3 expression level between the patients with acute myeloid leukemia and non-hematologic malignancies (P > 0.05), but the Pim-3 expression level was significantly lower in patients before chemotherapy as compared with that in patients post chemotherapy (P < 0.01). The comparison of Pim-3 expression before and after chemotherapy in remission group showed that Pim-3 expression levels before chemotherapy were all significantly lower than those after chemotherapy (P < 0.01), but there were no significant differences of Pim-3 expression levels between patients before and after chemotherapy in non-remission group (P > 0.05). The Pim-3 expression levels of non-remission patients after chemotherapy were all significantly lower than those of the remission patients after chemotherapy (P < 0.01). CONCLUSION: Pim-3 gene is abnormally expressed in the AML patients before and after chemotherapy, and this gene may be involved in the genesis and development of acute myeloid leukemia.