| Objective:Mantle cell lymphoma belongs to refractory B cell non Hodgkin's lymphoma with an aggressive clinical course and a poor prognosis,currently lacks effective therapies.Decitabine(DAC)is a kind of demethylation drugs with an obviously antitumor activity.At present,there is a consensus on the treatment of myelodysplastic syndrome(MDS)and refractory acute myeloid leukemia(AML),but there are few reports of its treatment for MCL.The proteasome inhibitor-Brotezomib(BTZ)has been approved for the treatment of patients with multiple myeloma.In recent years,many studies have demonstrated that bortezomib also play an impotant role in the treatment of patients with relapsed or refractory MCL and diffuse large B cell lymphoma,NCCN has been recommended that Bortezomib is only used as second-line therapy for mantle cell lymphoma.In this study,we investigated the effect of DAC combined with BTZ on proliferation and apoptosis of human MCL cell line and its mechanism of action,so as to provide experimental basis for the combination treatment in patients with MCL.Methods:1 Jeko-1 and Grante519 cells were treated with different concentrations of DAC and BTZ in vitro;Detecting the effect of drugs on the proliferation of MCL cell line by CCK8 method.2 Detecting the effect of drugs on the apoptosis rate by flow cytometry(FCM).3 RT-PCR was used to detect the m RNA levels of PCDH8,NF-Kb,Caspas3,Bcl-2,CCND1,Bax genes.4 Western blot was used to detect the protein levels of PCDH8,NF-Kb(P65),Caspas3,Bcl-2,CCND1,Bax genes.Results:1 Low concentrations of DAC(0.1,1,10?mol/L)could significantly inhibit the proliferation and induce apoptosis of Jeko-1 and Grante519 cells,the inhibitory effect was does and time dependent;We observes that DAC increased the expression level of PCDH8,Caspase3 and Bax,decreased the expression level of Bcl-2,CCND1 of the Jeko-1 and Grante519 cells;but the expression level of NF-KB(P65)had no obvious effect;2 High concentrations of BTZ(0.2,0.4?mol/L)could significantly inhibit the proliferation and induce apoptosis of Jeko-1,Grante519 cells,the inhibitory effect was does and time dependent;BTZ can increased the expression of Caspase3,Bax,decreased the expression of NF-KB(P65),Bcl-2,but the expression of PCDH8 and CCND1 had no a significant effect;3 DAC combined with BTZ significantly increased the inhibition rate and apoptosis rate of Jeko-1 and Grante519 cells;Moreover,The efficiency in DAC is better than BTZ;Compared with DAC and BTZ single drug,Combination therapy could obviously increase the expression levels of PCDH8,Caspase3 and Bax,and the expression levels of NF-KB(P65),decreased the the expression levels of Bcl-2 and CCND1.Conclusion:1 DAC can inhibit proliferation and promote apoptosis of Jeko-1 and Grante519 cells.In a certain concentration range,it is time dependent and dose dependent;The mechanism for DAC induced-antitumor activity may be the demethylation of DAC;Moreover,DAC could induce the increase of Caspase3 and Bax expression,decrease the expression level of Bcl-2 and CCND1,and further regulate the process of cell proliferation and apoptosis.2 BTZ can enhance the proliferation inhibition and apoptosis of Jeko-1 and Grante519 cells in a certain concentration range;The mechanism may be associated with BTZ can reduce the activity of NF-KB signaling pathway,leading to increasing expression of Caspase3,and Bax,decreasing the expression level of Bcl-2,so as to promote apoptosis and inhibit cell proliferation.3 Low concentration of DAC has demethylation,and high concentration of DAC has cytotoxicity.4 DAC and BTZ has a synergistic effect.The possible mechanism is the combination of two drugs enhanced the inhibition of the NF-KB signal pathway,and down regulation the level of Bcl-2,up regulation the level of Bax,which caused the expression level of Caspase3 protein increased further,to achieve higher antitumor effect.This conclusion provides a new theoretical basis for clinical treatment in the future. |
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