硼替佐米和来那度胺对淋巴瘤细胞株SNK-1、SNK-6和SNT-8的细胞毒性及诱导凋亡作用研究

Study on effects of bortezomib and lenalidomide on the cytotoxicity and induced apoptosis on lymphoma cell line SNK-1,SNK-6 and SNT-8

目的研究硼替佐米和来那度胺对淋巴瘤细胞株SNK-1、SNK-6和SNT-8细胞毒性及诱导凋亡作用。方法采用不同浓度的硼替佐米(1 nmol/L、10 nmol/L、30 nmol/L、100 nmol/L)分别对NK/T淋巴瘤细胞株作用24 h。采用不同浓度的来那度胺(1μmol/L、10μmol/L、30μmol/L、100μmol/L)分别对NK/T淋巴瘤细胞株作用24 h。通过台盼蓝拒染法对NK/T淋巴瘤细胞进行计数并对细胞活力进行测定,另采用流式细胞术和Annexin V/PI法对细胞凋亡进行测定,同时采用流式细胞仪对细胞周期分布进行测定。结果随着硼替佐米浓度(1 nmol/L、10 nmol/L、30 nmol/L、100 nmol/L)的升高,细胞存活率明显降低(P <0. 01)。流式细胞术检测结果发现,硼替佐米(30 nmol/L)可有效诱导NK/T淋巴瘤细胞株SNK-1、SNK-6和SNT-8凋亡,并且随着硼替佐米作用时间的延长(0 h、24 h、48 h),三种细胞株凋亡率明显升高(P <0. 01)。随着来那度胺(1μmol/L、10μmol/L、30μmol/L、100μmol/L)浓度的升高,细胞存活率并无明显变化(P> 0. 05)。NK/T淋巴瘤细胞株分别采用来那度胺(浓度为10μmol/L)处理0 h、24 h、48 h后,发现SNK-1、SNK-6、SNT-8在G0/G1期、G2/M期的分布并无显著差异(P> 0. 05)。结论硼替佐米作为一种临床常用的具有选择性的蛋白酶体抑制剂,对NK/T淋巴瘤细胞具有显著的诱导凋亡作用,对NK/T淋巴瘤的治疗具有潜在应用价值,可考虑作为临床治疗NK/T淋巴瘤的重要药物。在体外实验中,来那度胺对NK/T淋巴瘤细胞株存活、凋亡并无明显影响,但有关其在体内能否具有抗肿瘤活性的作用,则需今后深入探讨。

Objective To study the effects of bortezomib and lenalidomide on the cytotoxicity and induced apoptosis of SNK-1,SNK-6 and SNT-8 cells in lymphoma cell lines.Methods NK/T lymphoma cell lines were treated with different concentrations of bortezomib(1 nmol/L,10 nmol/L,30 nmol/L,100 nmol/L)for 24 h.NK/T lymphoma cell lines were treated with different concentrations of lenalidomide(1μmol/L,10μmol/L,30μmol/L,100μmol/L)for 24 h.NK/T lymphoma cells were counted by trypan blue exclusion method,and cell viability was measured.The apoptosis was detected by flow cytometry and Annexin V/PI.Meanwhile,the cell cycle distribution was detected by flow cytometry.Results Different concentrations of bortezomib(1 nmol/L,10 nmol/L,30 nmol/L,100 nmol/L)were used to treat NK/T lymphoma cell line for 24 h.It was found that cell viability was significantly decreased with increasing concentration(P<0.01).The results of flow cytometry showed that bortezomib(30 nmol/L)could effectively induce the apoptosis of NK/T lymphoma cell lines SNK-1,SNK-6 and SNT-8,and the apoptosis rate of the three cell lines increased significantly with the increasing time(P<0.01).The effects of different concentrations of lenalidomide(1μmol/L,10μmol/L,30μmol/L,100μmol/L)on NK/T lymphoma cell line were 24 h,respectively.It was found that cell viability did not change significantly with increasing concentration(P>0.05).When the NK/T lymphoma cells were treated with lenalidomide(10μmol/L)for 0 h,24 h and 48 h,it was found that there was no significant difference in the distribution of SNK-1,SNK-6 and SNT-8 during G 0/G 1 and G 2/M stage(P>0.05).Conclusion Bortezomib as a clinical common selective proteasome inhibitor,has a significant effect on the NK/T induced apoptosis of lymphoma cells,which has potential application to treat NK/T lymphoma,so it can be considered as an important drug in clinical treatment of NK/T lymphoma.In vitro experiments,lenalidomide has no significant effect on the survival and apoptosis of NK/T lymphoma cell lines,but whether it can have antitumor activity in vivo needs further study.