摘要
目的探讨蛋白酶体抑制剂硼替佐米(bortezomib,BZM)增强Vγ9Vδ2T细胞对套细胞淋巴瘤的杀伤作用及其相关机制。方法通过帕米磷酸二钠和IL-2刺激人外周血中单个核细胞得到Vγ9Vδ2T细胞。给予BZM刺激后,溴乙啡锭二聚体-1(ethidium homodimer-1,EthD-1)染色法检测Vγ9Vδ2T细胞对Maver细胞的杀伤效应;CD107a标记法检测Vγ9Vδ2T细胞的脱颗粒效应;CD25与CD69标记法检测Vγ9Vδ2T细胞的活化状况;ELISA法检测Vγ9Vδ2T细胞的TNF-α和IFN-γ的生成水平;Western blot检测Vγ9Vδ2T细胞中Notch1和p-NF-κB水平。结果 BZM增强Vγ9Vδ2T细胞对Maver细胞的杀伤效应和增加Vγ9Vδ2T细胞的脱颗粒能力(CD107a表达增加)。BZM增强Vγ9Vδ2T细胞活化(CD25与CD69的表达增加)以及活化效应功能(TNF-α和IFN-γ的生成水平增加)。BZM能增强Vγ9Vδ2T细胞的Notch1/NF-κB信号(Notch1和p-NF-κB水平增加)。给予Notch1和NF-κB抑制剂处理Vγ9Vδ2T细胞,BZM的以上效应可被部分逆转。结论 BZM可通过Notch/NF-κB通路增强Vγ9Vδ2T细胞的活化,并进一步增强Vγ9Vδ2T细胞对Maver细胞的杀伤效应。
Objective To explore the enhancement of the tumor killing effect of Vγ9Vδ2T cells by proteasome inhibitor bortezomib(BZM) on mantle cell lymphoma(MCL) and its underlying mechanisms. Methods Vγ9Vδ2T cells were obtained by stimulating human peripheral blood mononuclear cells with pamidronate disodium and IL-2. After BZM stimulation, the killing effect of Vγ9Vδ2T cells on Maver cells was detected by ethidium homodimer-1(EthD-1) staining;the degranulation effect of Vγ9Vδ2T cells was detected by CD107 a labeling method;the activation of Vγ9Vδ2T cells was detected by CD25 and CD69 labeling method;the production levels of TNF-α and IFN-γ in Vγ9Vδ2T cells were detected by ELISA;the expression levels of Notch1 and p-NF-κB in Vγ9Vδ2T activated cells were examined by Western blotting. Results BZM enhanced the killing effect of Vγ9Vδ2T cells on Maver cells and improved the degranulation capacity of Vγ9Vδ2T cells(the expression of CD107 a was increased). BZM promoted the activation of Vγ9Vδ2T cells(the expressions of CD25 and CD69 were increased) and the subsequent activation effect function(the productions of TNF-α and IFN-γ were increased). BZM enhanced the Notch1/NF-κB signaling pathway in Vγ9Vδ2T cells(the expressions of Notch1 and p-NF-κB were increased). Treating Vγ9Vδ2T cells with Notch1 and NF-κB inhibitors partially reversed the above effects of BZM. Conclusions BZM promotes the activation of Vγ9Vδ2T cells through Notch/NF-κB signaling pathway, and further enhances the killing effect of Vγ9Vδ2T cells on Maver cells.
作者
张岩
邓黎黎
张壮苗
Zhang Yan;Deng Lili;Zhang Zhuangmiao(Department of Hematology and Oncology,Sanya People’s Hospital,Sanya 572000,China)
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2020年第1期15-22,共8页
Chinese Journal of Histochemistry and Cytochemistry
基金
海南省三亚市科技创新局资助项目(2011YD88)。
