硼替佐米和来那度胺对NK/T淋巴瘤细胞作用的实验研究

Experimental study on effects of bortezomib and lenalidomide on NK/T-cell lymphoma cell lines

目的通过体外细胞培养观察硼替佐米和来那度胺对NK/T细胞淋巴瘤(NK/TCL)细胞株凋亡的作用,寻找治疗NK/T淋巴瘤的潜在药物。方法利用台盼蓝拒染法检测细胞活力及计数细胞,流式细胞术检测细胞凋亡和细胞周期分布,Annexin V/PI法检测细胞凋亡。结果硼替佐米对SNK-1、SNK-6、SNT-8有明显的细胞毒作用,在临床治疗浓度下显著诱导凋亡而减少细胞的存活。硼替佐米(30 nmol/L)作用24 h,SNK-1、SNK-6、SNT-8细胞存活比例分别为(47.2±3.5)%、(33.5±3.9)%、(52.1±4.3)%,与对照组相比差异均有统计学意义(P<0.01)。硼替佐米对3种细胞株活性抑制IC50的浓度分别为19.5、12.7、20.7 nmol/L。流式细胞术对细胞凋亡检测发现硼替佐米对SNK-1(30 nmol/L)、SNK-6(10 nmol/L)、SNT-8(30 nmol/L)细胞株均有显著的诱导凋亡作用,作用24 h后凋亡率为(47.0±4.2)%、(43.0±3.8)%、(53.0±4.1)%。来那度胺对3种细胞株的存活和增殖无显著作用,对细胞周期的分布也无明显影响。来那度胺联合硼替佐米无协同促进细胞凋亡作用。结论硼替佐米是选择性的蛋白酶体抑制剂,具有诱导NK/TCL细胞株凋亡的作用,可能有成为NK/TCL治疗药物的潜在价值。来那度胺在体外实验中对NK/TCL细胞株无显著作用,与硼替佐米亦无协同作用,体内是否具有抗肿瘤活性尚需进一步的研究。

Objective To investigate the effects of bortezomib and lenalidomide on the induction of cell apoptosis in natural killer / T-cell lymphoma（ NK / TCL） cell lines. Methods Trypan blue exclusion test was used to account living cells.Flow cytometry was applied to detect cell apoptosis and contents of cellular DNA for analysis of cell cycle distribution. The apoptotic cells were quantified by dual labeling of recombinant annexin V and propidium iodide. Results The cytotoxic effects of bortezomib on SNK-1,SNK-6 and SNT-8 were observed. Bortezomib at clinical therapeutic concentrations significantly reduced the cells survival by inducing cell apoptosis in NK / TCL cell lines. The percentages of survival cells were（ 47. 2 ± 3. 5） %,（ 33. 5 ± 3. 9） %,（ 52. 1 ± 4. 3） % for SNK-1,SNK-6,and SNT-8,respectively,after bortezomib（ 30 nmol / L） treatment for 24 hours（ P〈0. 01 as compared with control）. The IC 50 of bortezomib for SNK-1,SNK-6,and SNT-8 were 19. 5,12. 7,and 20. 7 nmol / L,respectively. By flow cytometry analysis,we found that 24 hours exposure to bortezomib significant induced apoptosis with（ 47. 0 ± 4. 2） % for SNK1（ 30 nmol / L）,43. 0 ± 3. 8% for SNK6（ 10 nmol /L）,and（ 53. 0 ± 4. 1） % for SNK-8（ 30 nmol / L）. Lenalidomide showed no significant effect on cell survival,cell proliferation and distribution of cell cycle. Lenalidomide in combination with bortezomib showed no synergistic effect on inducing cell apoptosis. Conclusion Bortezomib is a powerful agent in inducing NK / TCL cells apoptosis. It is a potential therapeutic agent in NK / TCL treatment. Lenalidomide showed no significant effect on NK / TCL cells in culture; moreover,it showed no synergistic effect with bortezomib on inducing cell apoptosis. Whether lenalidomide has effect on NK / TCL in vivo should be further studied.