Chidamide triggers pyroptosis in T-cell lymphoblastic lymphoma/leukemia via the FOXO1/GSDME axis.

BACKGROUND: T-cell lymphoblastic lymphoma/leukemia (T-LBL/ALL) is an aggressive form of hematological malignancy associated with poor prognosis in adult patients. Histone deacetylases (HDACs) are aberrantly expressed in T-LBL/ALL and are considered potential therapeutic targets. Here, we investigated the antitumor effect of a novel HDAC inhibitor, chidamide, on T-LBL/ALL. METHODS: HDAC1, HDAC2 and HDAC3 levels in T-LBL/ALL cell lines and patient samples were compared with those in normal controls. Flow cytometry, transmission electron microscopy and lactate dehydrogenase release assays were conducted in Jurkat and MOLT-4 cells to assess apoptosis and pyroptosis. A specific forkhead box O1 (FOXO1) inhibitor was used to rescue pyroptosis and upregulated gasdermin E (GSDME) expression caused by chidamide treatment. The role of the FOXO1 transcription factor was evaluated by dual-luciferase reporter and chromatin immunoprecipitation assays. The efficacy of chidamide in vivo was evaluated in a xenograft mouse. RESULTS: The expression of HDAC1, HDAC2 and HDAC3 was significantly upregulated in T-LBL/ALL. Cell viability was obviously inhibited after chidamide treatment. Pyroptosis, characterized by cell swelling, pore formation on the plasma membrane and lactate dehydrogenase leakage, was identified as a new mechanism of chidamide treatment. Chidamide triggered pyroptosis through caspase 3 activation and GSDME transcriptional upregulation. Chromatin immunoprecipitation assays confirmed that chidamide led to the increased transcription of GSDME through a more relaxed chromatin structure at the promoter and the upregulation of FOXO1 expression. Moreover, we identified the therapeutic effect of chidamide in vivo. CONCLUSIONS: Our study suggested that chidamide exerts an antitumor effect on T-LBL/ALL and promotes a more inflammatory form of cell death via the FOXO1/GSDME axis, which provides a novel choice of targeted therapy for patients with T-LBL/ALL.

A novel prognostic nomogram based on imaging and molecular parameters for newly diagnosed extranodal natural killer/T-cell lymphoma patients.

Not available.

Optimized BEAC conditioning regimen improves clinical outcomes of autologous hematopoietic stem cell transplantation in non-Hodgkin lymphomas.

The conditioning regimen is an important part of autologous hematopoietic stem cell transplantation (ASCT). We explored the efficacy and safety of an optimized BEAC (adjusted-dose, intermediate-dose cytarabine and reduced-dose cyclophosphamide, AD-BEAC) conditioning regimen for non-Hodgkin lymphoma (NHL). A total of 141 NHL patients received AD-BEAC or a standard-dose BEAC (SD-BEAC) conditioning regimen from January 2007 to December 2017, and 104 patients were included in the study after 1:1 propensity matching. The 5-year overall survival (OS) and progression free survival (PFS) rates were significantly higher with AD-BEAC than with SD-BEAC (82.7% vs. 67.3%, P = 0.039; 76.9% vs. 57.7%, P = 0.039). Transplant-related mortality (TRM) was 3.8% in both the AD-BEAC and SD-BEAC groups. The AD-BEAC group had lower incidence of oral ulcers and cardiotoxicity than the SD-BEAC group. An optimized BEAC conditioning regimen is an effective conditioning regimen for ASCT in NHL with acceptable toxicity, that is more effective and safer than a standard BEAC conditioning regimen.

[Effects of Marsdenia tenacissima extract on proliferation and apoptosis of hematologic neoplasm cell line cells].

OBJECTIVE: To investigate the effects of the extract from Marsdensia tenacissima on proliferation and apoptosis of human hematologic neoplasm cell line cells. METHODS: Raji, NB4 and K562 cells were treated in vitro with different concentrations of the extract from Marsdensia tenacissima, including different ethanol elution components and C21 steroidal saponin monomer compounds, for different periods. Tumor cell proliferation was measured by MTT colorimetric assay and its apoptosis was determined by the flow cytometry (FCM). RESULTS: Firstly, with higher concentrations, 100 microg/mL and 200 microg/mL, 70% ethanol eluate from Marsdensia tenacissima inhibited the proliferation of Raji, NB4 and K562 cells significantly, in a dose and time dependent manner, compared with 30% and 50% ethanol elution components from Marsdensia tenacissima (P < 0.05). Secondly, four C21 steroidal saponin monomer compounds, tenacissosides B,C,I and marsdenoside K, also inhibited the proliferation of Raji, NB4 and K562 cells in vitro significantly, in a dose and time dependent manner, compared with that of control group (P < 0.05). Among them, tenacissoside C showed the strongest inhibition effects on proliferation of these cells under all experimental conditions compared with the other three C21 steroidal saponin monomer compounds (P < 0.05). Furthermor, the IC50 of tenacissosides C on proliferation of Raji, NB4 and K562 cells were 64.1 micromol/L, 70.4 micromol/L and 105.8 micromol/L, respectively. Finally, after Raji, NB4 and K562 cells were treated with 98.4 micromol/L tenacissoside C for 24 h and 48 h, the early apoptosis rates and late apoptosis rates of these tumor cells increased markedly, compared with the control group (P < 0.05). CONCLUSION: The extract from Marsdensia tenacissima, including different ethanol elution components and C21 steroidal saponin monomer compounds, may inhibit the proliferation of some human hematologic neoplasm cell line cells and induce these tumor cells apoptosis in vitro, especially tenacissoside C, one of the C21 steroidal saponin monomer compounds, showed the strongest effects on proliferation of these tumor cells when compared with other ones, with the strongest inhibition activities on human Burkitt's lymphoma cell line Raji cells.

[Antitumor effects of matrix protein of vesicular stomatic virus on EL4 lymphoma mice].

OBJECTIVE: To explore antitumor effects of plasmid pcDNA3. 1-MP encoding matrix protein of vesicular stomatitis virus (VSV) complexed with cationic liposome (DOTAP:CHOL) in mice with EL4 lymphoma. METHODS: C57BL/6 mouse model with EL4 lymphoma was established. Sixty mice bearing EL4 lymphoma were divided randomly into five groups including Lip-MP, Lip-pVAX, Lip, ADM and NS groups, which were intravenously injected with liposome-pcDNA 3. 1-MP complex, liposome-pVAX complex, empty liposome, Adriamycin and normal saline respectively every three days. Tumor volumes and survival time were monitored. Microvessel density and tumor proliferative index in tumor tissues were determined by CD31, Ki-67 immunohistochemistry staining, meanwhile the tumor apoptosis index was measured by TUNEL method. RESULTS: From 6 days after treatments on, the tumor volume in Lip-MP group was much smaller than that in Lip-pVAX, Lip and NS group (P < 0.05). The median survival time of mice in Lip-MP group, 44 days after inoculation of tumor cells, was significantly higher than that in other groups (P < 0.05), which was 39 days, 38.5 days and 34 days in Lip-pVAX, Lip and NS groups respectively. The MVD value in tumor tissues in Lip-MP group was less than that in Lip-pVAX, Lip and NS groups (P < 0.05). Ki67 staining revealed that Lip-MP complex apparently suppressed the proliferation of EL4 tumor cells in vivo (P < 0.05). TUNEL assays showed that apoptosis index of tumor cells in Lip-MP group, 10.60 +/- 1.71, was much higher than that in other three groups (P < 0.05). CONCLUSIONS: Lip-MP complex, the plasmid encoding matrix protein of VSV (VSV-MP) encapsulated in cationic liposome, significantly inhibited the growth of tumor and prolonged the survival of mice bearing EL4 lymphoma, which may be related to the induction of tumor cell apoptosis, inhibition of tumor angiogenesis, and suppression of tumor cell proliferation.

[Gene cloning of mIFN-gamma and construction of recombinant adenovirus].

OBJECTIVE: To develop a rapid and efficient method for preparing recombinant adenovirus containing mouse IFN-gamma (mIFN-gamma) gene by homologous recombination in E. coli. in order to build a foundation for research into gene therapy of liver fibrosis. METHODS: The target gene mIFN-gamma was amplified by using PCR from the vector pORF5-mIFN-gamma. Once verified, it was cut out by double endonucleases, then connected to the shuttle vector pAdTrack-CMV. The newly constructed vector was linearized by Pme I following transformation to the E. coli. BJ5183, which contained the backbone vector pAdEasy-1. The correct recombinant pAd-mIFN-gamma was selected by endonucleases and by Kanamycin resistance. Again it was linearized with Pac I , then transfected to AD-293 cells by means of Calcium Phosphate method. Finally, the target gene IFN-gamma was identified by PCR and Western blot methods. RESULTS: The target gene mIFN-gamma amplified by PCR was identified by DNA sequencing, which proved that the mIFN-gamma gene consisted of 468 nucleotides and was completely the same with the sequence published on the GenBank. The adenoviral vector constructed by homologous recombination had the gene of interest and the viral could be examined 4-6 days after transfection, and the green fluorescence intensity became greater at about 8-11 days. The adenovirus obtained at the 12th day was digested by protease K and then was amplified by PCR and identified by Western blot. The two methods proved that the adenovirus encoded the target gene mIFN-gamma. CONCLUSION: Preparing recombinant adenovirus containing mIFN-gamma gene by homologous recombination in E. coli. Is a rapid and efficient method. The Ad-mIFN-gamma can be propagated in 293 cell line. It may be used as a novel agent for gene therapy in liver fibrosis.