The Molecular Mechanisms Of Glycolytic Pathway In Malignant Biological Behaviors Of Lymphoma And Targeted Intervention Study

Objective:To investigate the effects of glycolysis-related gene-silencing by shRNA interference on the malignant biological behaviors of human lymphoma cell lines,and the mechanisms of rituximab?RTX?in combination with bortezomib?BTZ?for aggressive B-cell lymphoma through glycolytic signaling pathway,therefore to provide theoretical basis for optimizing lymphoma clinical therapeutical scheme.Methods:The stable-transfected lymphoma cell lines were constructed with lentiviral vectors carried shRNAs targeting glycolysis-related genes,including HIF-1?,HK II and c-myc.The expression levels of mRNA and protein were detected by RT-qPCR and Western blotting,respectively.The cell viability was assessed using trypan blue staining,and the cell growth curve was drawn.The IC₅₀0 of doxorubicin?DOX?for lymphoma cells was examined by CCK-8 method.The cell cycle distribution and apoptosis were analyzed by flow cytometry.The concentrations of lactic acid and glucose in the supernatant of cell culture were analyzed using Lactic Acid Detection Kit and Glucose?HK?Assay Kit,respectively.Results:The lymphoma cell lines transfected with lentiviral vectors carried with shRNAs targeting glycolysis-related genes,including HIF-1?,HK II and c-myc,were established,while the control vectors carried with non-mammalian shRNAs were used as the control groups.It is confirmed that the aforementioned glycolysis-related genes participate in regulation of the malignant biological behaviors of lymphoma cells.After lentivirus-mediated transduction with shRNA targeting glycolysis-related genes,the proliferation of lymphoma cells was significantly inhibited,the cell cycle was arrested and the cell apoptosis was induced.Gene-silencing also reversed the aberrant metabolic phenotype of lymphoma cells through the inhibition of glucose consumption and lactic acid generation,and enhanced cell sensitivity to anticancer drugs,therefore attenuated the malignant biological behaviors of lymphoma cells.Apoptosis of lymphoma cell lines was induced by the down-regulation of Bcl-2protein and the up-regulation of caspase-3 cleavage.Rituximab in combination with bortezomib could induce cell apoptosis,arrest cell cycles and hamper the glucose consumption and lactic acid generation of lymphoma cells via down-regulating the transcriptional levels of glycolysis-related genes,including HIF-1?,HK II and LDHA,therefore play an efficient role against lymphoma.Conclusion:It suggested that glycolysis-related genes,including HIF-1?,HK II and c-myc,may be the potential targets for the treatment of aggressive B-cell lymphoma,and targeted interference could be a novel promising approach for refractory/relapsed aggressive lymphomas.The combination of rituximab with bortezomib is prospective to become a new strategy for the theratment of aggressive B-cell lymphoma and also provide novel foundation for its anti-lymphoma mechanism in glycolytic pathway.