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
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摘要
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,10 μmol/L,30 μmol/L,100 μmol/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( 1 μmol/L,10 μmol/L,30 μmol/L,100 μmol/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( 10 μmol/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 G0/G1 and G2/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.
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,10 μmol/L,30 μmol/L,100 μmol/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( 1 μmol/L,10 μmol/L,30 μmol/L,100 μmol/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( 10 μmol/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 G0/G1 and G2/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.
引文
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[11]华佳叶,周旭红,欧阳舒婷,等.硼替佐米对伊马替尼耐药细胞株K562的凋亡作用及机制[J].南方医科大学学报,2017,37(8):1136-1139.
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[14]王莹莹,钟瑶,俞夜花,等.硼替佐米联合毛喉素诱导硼替佐米耐药骨髓瘤细胞凋亡的实验研究[J].中国癌症杂志,2016,26(9):784-789.
[15]郑军,范松青.靶向抗肿瘤小分子化合物类药物的研究[J].国际肿瘤学杂志,2012,39(3):172-175.
[2]杨融辉,李诗文,沈静,等.来那度胺治疗弥漫大B细胞淋巴瘤的进展[J].现代肿瘤医学,2018,26(1):120-123.
[3]王江涛,关涛,苏丽萍,等.淋巴瘤的免疫治疗[J].国际肿瘤学杂志,2017,44(8):633-637.
[4] Mourtada-Maarabouni M,Williams GT. Role of GAS5 noncoding RNA in mediating the effects of rapamycin and its analogues on mantle cell lymphoma cells[J]. Clin Lymphoma Myeloma Leuk,2014,14(6):468-473.
[5]周静静,冯献启,聂淑敏,等.免疫表型对以硼替佐米为主方案治疗的多发性骨髓瘤患者预后的影响[J].白血病·淋巴瘤,2018,27(8):459-463.
[6]姜立彩,罗建民.硼替佐米对淋巴瘤细胞增殖、凋亡及SHP-2基因表达的影响[J].中国实验血液学杂志,2015,23(4):1026-1029.
[7] Yoshimura M,Ishizawa J,Ruvolo V,et al. Induction of p53-mediated transcription and apoptosis by exportin-1(XPO1)inhibition in mantle cell lymphoma[J]. Cancer Sci,2014,105(7):795-801.
[8]杜超,翁巍,孙致信,等.硼替佐米和来那度胺对NK/T淋巴瘤细胞作用的实验研究[J].中华全科医学,2016,14(3):377-380.
[9]金玉龙,徐莉,唐海龙,等.丙戊酸钠协同硼替佐米对RPMI8226细胞的促凋亡作用[J].现代生物医学进展,2015,15(14):2649-2651,2657.
[10] Sekihara K,Saitoh K,Han L,et al. Targeting mantle cell lymphoma metabolism and survival through simultaneous blockade of m TOR and nuclear transporter exportin-1[J]. Oncotarget,2017,8(21):34552-34564.
[11]华佳叶,周旭红,欧阳舒婷,等.硼替佐米对伊马替尼耐药细胞株K562的凋亡作用及机制[J].南方医科大学学报,2017,37(8):1136-1139.
[12] Dunn JP. Interleukins in the Diagnosis of Intraocular Lymphoma:Do We Still Need Histologic Confirmation[J]? Retina,2018,38(4):647-649.
[13] Hanauer JDS,Rengstl B,Kleinlützum D,et al. CD30-targeted oncolytic viruses as novel therapeutic approach against classical Hodgkin lymphoma[J]. Oncotarget,2018,9(16):12971-12981.
[14]王莹莹,钟瑶,俞夜花,等.硼替佐米联合毛喉素诱导硼替佐米耐药骨髓瘤细胞凋亡的实验研究[J].中国癌症杂志,2016,26(9):784-789.
[15]郑军,范松青.靶向抗肿瘤小分子化合物类药物的研究[J].国际肿瘤学杂志,2012,39(3):172-175.