Contributions
Abstract: PB1625
Type: Publication Only
Background
T cell acute lymphoblastic leukemia (T-ALL) is a hematopoietic clonal malignancy caused by the malignant transformation of T lymphocyte driven by gene mutation. The prognosis of T-ALL is poor and early relapse is common.
Aims
We aimed at looking for specific and effective therapeutic target for T-ALL and eventually cure this form of leukemia by targeted therapy.
Methods
Bone marrow mononuclear cells (BMMC) are collected from bone marrow samples of T-ALL patients, including at initial presentation (n=46), during first CR (n=23) and at relapse (n=6). The expression level of mRNA encoding L-cellular Fas-associated death domain-like interleukin-1β converting enzyme inhibitory protein (c-FLIP) was assessed by realt time PCR. Changes in the expression level of HDAC before and after chidamide treatment were also assessed by western blot. Necroptosis and apoptosis after chidamide treatment were assessed by flow cytometry. Changes in expression level of c-FLIPL protein before and after treatment were assessed by western blot. Expression level of early apoptotic protein, key proteins of necroptosis were assessed by western blot.The effect of chidamide on NF-κB signaling pathway activity and expression of key molecules when inducing necroptosis were assessed by western blot. The regulating effect of chidamide on downstream genes of NF-κB pathway including cyclinD1, TNFα, lL-2, IL-8 were assessed by real- time PCR.
Results
The expression level of c-FLIPL mRNA is significantly higher in patients at initial presentation and relapse, compared to those at complete remission and healthy control. The expression level of c-FLIPL mRNA is associated with patient risk stratification, white blood cell count at initial presentation, serum level of lactate dehydrogenase (LDH), serum level of hydroxybutyrate dehydrogenase (HBDH), CD45, HLA-DR, SIL-TAL1 fusion gene and complex karyotype, and is not associated with age, sex, plasma fibrinogen level, and the chromosomal aberration 6q-. Patients who did not achieve CR during first chemotherapy had a higher c-FLIPL mRNA level than those who did (p<0.05).The expression level of histone deacetylase is higher in bone marrow mononuclear cells of T-ALL patients, Jurkat and HUT-78 cell lines. After treatment with chidamide, the expression level of histone deacetylase was significantly decreased in both cell lines.Chidamide induced necroptosis and apoptosis in Jurkat and HUT-78 cell lines. After apoptosis inhibitor was applied, chidamide mainly exert its effect of inducing cell death by inducing necroptosis. Chidamide inhibits the transduction and translation to c-FLIPL gene. When apoptosis is inhibited, chidamide upregulates the expression level of receptor-interacting protein 3 (RIP3) and the phosphorylation level of mixed lineage kinase domain-like (MLKL). After treatment with chidamide, the phosphorylation level of I-κB and p65 protein were both significantly decreased.
Conclusion
c-FLIPL mRNA expression level is abnormally high in T-ALL patients both at initial presentation and at relapse. The expression level of c-FLIPL is associated with risk stratification, white blood cell count, serum LDH level, serum HBDH level, CD45, SIL-TAL1 fusion gene, complex karyotype and disease outcome. c-FLIPL could be used as a prognostic marker in T-ALL.Chidamide suppresses histone deacetylation in Jurkat and HUT-78 cell lines.Chidamide induces necroptosis in Jurkat and HUT-78 cell lines by down regulating the transcription and translation of c-FLIPL gene.Chidamide induces necroptosis in Jurkat and HUT-78 cell lines via the classical NF-κB signaling pathway.
Session topic: 1. Acute lymphoblastic leukemia - Biology
Keyword(s): Acute lymphoblastic leukemia
Abstract: PB1625
Type: Publication Only
Background
T cell acute lymphoblastic leukemia (T-ALL) is a hematopoietic clonal malignancy caused by the malignant transformation of T lymphocyte driven by gene mutation. The prognosis of T-ALL is poor and early relapse is common.
Aims
We aimed at looking for specific and effective therapeutic target for T-ALL and eventually cure this form of leukemia by targeted therapy.
Methods
Bone marrow mononuclear cells (BMMC) are collected from bone marrow samples of T-ALL patients, including at initial presentation (n=46), during first CR (n=23) and at relapse (n=6). The expression level of mRNA encoding L-cellular Fas-associated death domain-like interleukin-1β converting enzyme inhibitory protein (c-FLIP) was assessed by realt time PCR. Changes in the expression level of HDAC before and after chidamide treatment were also assessed by western blot. Necroptosis and apoptosis after chidamide treatment were assessed by flow cytometry. Changes in expression level of c-FLIPL protein before and after treatment were assessed by western blot. Expression level of early apoptotic protein, key proteins of necroptosis were assessed by western blot.The effect of chidamide on NF-κB signaling pathway activity and expression of key molecules when inducing necroptosis were assessed by western blot. The regulating effect of chidamide on downstream genes of NF-κB pathway including cyclinD1, TNFα, lL-2, IL-8 were assessed by real- time PCR.
Results
The expression level of c-FLIPL mRNA is significantly higher in patients at initial presentation and relapse, compared to those at complete remission and healthy control. The expression level of c-FLIPL mRNA is associated with patient risk stratification, white blood cell count at initial presentation, serum level of lactate dehydrogenase (LDH), serum level of hydroxybutyrate dehydrogenase (HBDH), CD45, HLA-DR, SIL-TAL1 fusion gene and complex karyotype, and is not associated with age, sex, plasma fibrinogen level, and the chromosomal aberration 6q-. Patients who did not achieve CR during first chemotherapy had a higher c-FLIPL mRNA level than those who did (p<0.05).The expression level of histone deacetylase is higher in bone marrow mononuclear cells of T-ALL patients, Jurkat and HUT-78 cell lines. After treatment with chidamide, the expression level of histone deacetylase was significantly decreased in both cell lines.Chidamide induced necroptosis and apoptosis in Jurkat and HUT-78 cell lines. After apoptosis inhibitor was applied, chidamide mainly exert its effect of inducing cell death by inducing necroptosis. Chidamide inhibits the transduction and translation to c-FLIPL gene. When apoptosis is inhibited, chidamide upregulates the expression level of receptor-interacting protein 3 (RIP3) and the phosphorylation level of mixed lineage kinase domain-like (MLKL). After treatment with chidamide, the phosphorylation level of I-κB and p65 protein were both significantly decreased.
Conclusion
c-FLIPL mRNA expression level is abnormally high in T-ALL patients both at initial presentation and at relapse. The expression level of c-FLIPL is associated with risk stratification, white blood cell count, serum LDH level, serum HBDH level, CD45, SIL-TAL1 fusion gene, complex karyotype and disease outcome. c-FLIPL could be used as a prognostic marker in T-ALL.Chidamide suppresses histone deacetylation in Jurkat and HUT-78 cell lines.Chidamide induces necroptosis in Jurkat and HUT-78 cell lines by down regulating the transcription and translation of c-FLIPL gene.Chidamide induces necroptosis in Jurkat and HUT-78 cell lines via the classical NF-κB signaling pathway.
Session topic: 1. Acute lymphoblastic leukemia - Biology
Keyword(s): Acute lymphoblastic leukemia