柔红霉素对人B细胞淋巴瘤OCI-LY7细胞凋亡的诱导作用及其机制

doi:10.13481/j.1671-587x.20190625

摘要/Abstract

摘要： 目的：探讨柔红霉素对人B细胞淋巴瘤OCI-LY7细胞凋亡的影响及其机制，阐明柔红霉素在弥漫大B细胞淋巴瘤中的耐药机制。方法：将体外培养的OCI-LY7细胞分为0、0.1、1.0和10.0 mg·L^-1柔红霉素组。采用MTT法检测各组OCI-LY7细胞存活率，流式细胞术检测各组OCI-LY7细胞凋亡率，Western blotting法检测各组OCI-LY7细胞中凋亡相关蛋白Bax、cleaved caspase-3和survivin表达水平。后续实验分为对照组、柔红霉素组、柔红霉素+空载体组和柔红霉素+survivin组，按照上述方法检测survivin过表达后各组OCI-LY7细胞存活率、凋亡率和细胞中凋亡相关蛋白表达水平。结果：与0 mg·L^-1组比较，处理48 h后0.1、1.0和10.0 mg·L^-1柔红霉素组OCI-LY7细胞存活率和细胞中survivin蛋白表达水平逐渐降低（P<0.05），而细胞凋亡率和细胞中cleaved caspase-3及Bax蛋白表达水平逐渐升高（P<0.05）。与对照组比较，柔红霉素组OCI-LY7细胞存活率和细胞中survivin蛋白表达水平明显降低（P<0.05），而细胞凋亡率和细胞中cleaved caspase-3及Bax蛋白表达水平明显升高（P<0.05）；与柔红霉素组比较，柔红霉素+survivin组OCI-LY7细胞存活率和细胞中survivin蛋白表达水平明显升高（P<0.05），而细胞凋亡率和细胞中cleaved caspase-3及Bax蛋白表达水平明显降低（P<0.05）。结论：柔红霉素可诱导OCI-LY7细胞凋亡，其作用机制可能与下调survivin蛋白表达有关。

关键词: 人B细胞淋巴瘤, 柔红霉素, 细胞凋亡, survivin

Abstract: Objective: To investigate the effect of daunorubicin on the apoptosis of human B-cell lymphoma OCI-LY7 cells and its mechanism,and to elucidate the drug resistance mechanism of daunorubicin in the diffuse large B-cell lymphoma. Methods: The OCI-LY7 cells cultured in vitro were divided into 0,0.1,1.0 and 10.0 mg·L^-1 daunorubicin groups.The survival rates of OCI-LY7 cells in various groups were detected by MTT assay,the apoptotic rates of the OCI-LY7 cells in various groups were detected by flow cytometry,and the expression levels of apoptosis-related proteins Bax,cleaved caspase-3, and survivin in the OCI-LY7 cells in various groups were detected by Western blotting method. The follow-up experiments were divided into control group,daunorubicin group,daunorubicin+vector group and daunorubicin+survivin group; the survival rates,the apoptotic rates and the expression levels of apoptosis-related proteins in the OCI-LY7 cells in various groups after overexpression of survivin were detected by the above-mentioned methods. Results: Compared with 0 mg·L^-1 group,the survival rates of OCI-LY7 cells and the expression levels of survivin protein in the OCI-LY7 cells in 0.1,1.0 and 10.0 mg·L^-1 daunorubicin groups were decreased gradually after treated for 48 h(P<0.05),while the apoptotic rates and the expression levels of Cleaved caspase-3 and Bax proteins in the OCI-LY7 cells were gradually increased (P<0.05). Compared with control group,the survival rate and the expression level of survivin protein in the OCI-LY7 cells in daunorubicin group were significantly decreased(P<0.05),while the apoptotic rate and the expression levels of cleaved caspase-3 and Bax proteins in the OCI-LY7 cells were significantly increased (P<0.05); compared with daunorubicin group,the survival rate and the expression level of survivin protein in the OCI-LY7 cells in daunorubicin+survivin group were significantly increased(P<0.05),while the apoptotic rate and the expression levels of cleaved caspase-3 and Bax protein in the OCI-LY7 cells in daunorubicin+vector group were significantly decreased (P<0.05). Conclusion: Daunorubicin can induce the apoptosis of OCI-LY7 cells,and its mechanism may be related to down-regulation of the expression of survivin protein.

Key words: human B cell lymphoma, daunorubicin, apoptosis, survivin

中图分类号:

R733.4

田鹏飞, 陈娟娟, 尹俊杰, 李冰, 郭珊. 柔红霉素对人B细胞淋巴瘤OCI-LY7细胞凋亡的诱导作用及其机制[J]. 吉林大学学报(医学版), 2019, 45(06): 1340-1345.

TIAN Pengfei, CHEN Juanjuan, YIN Junjie, LI Bing, GUO Shan. Inductive effect of daunorubicin on apoptosis of human B-cell lymphoma OCI-LY7 cells and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1340-1345.

参考文献

[1] 王猛,周晓莹,王立新.超声造影在非霍奇金淋巴瘤中的应用进展[J].临床与病理杂志,2018,38(9):2030-2033.
[2] SCHMITZ R,WRIGHT G W,HUANG D W,et al.Genetics and pathogenesis of diffuse large B-cell lymphoma [J].New Engl J Med,2018,378(15):1396-1407.
[3] 刘靖,刘芳,刘连,等.经典Wnt信号通路与弥漫大B细胞淋巴瘤[J].佛山科学技术学院学报:自然科学版,2015,33(4):1-4.
[4] MIZUNO S,HANAMURAI,OTAA,et al.Establishment and characterization of a novel vincristine‐resistant diffuse large B-cell lymphoma cell line containing the 8q24 homogeneously staining region [J].FEBS Open Biol,2018,8(12):1977-1991.
[5] GUTSCH J,WERTHMANN P G,ROSENWALD A,et al.Complete remission and long-term survival of a patient with a diffuse large B-cell lymphoma under viscum album extracts after resistance to R-CHOP: a case report [J].Anticancer Res,2018,38(9):5363-5369.
[6] YANG L,JIAN G E,XIA R.Resistance to daunorubicin in acute myeloid leukemia is associated with mutations in the TET2 gene [J].Acta Univer Med Anhui,2018,53(2):246-250.
[7] LIU YH,LEI PC,QIAO H,et al.MiR-9 Enhances the chemosensitivity of AML cells to daunorubicin by targeting the EIF5A2/MCL-1 axis[J].Int J Biol Scil,2019,15(3):579-586.
[8] WANG M,CAI W R,MENG R,et al.miR-485-5p suppresses breast cancer progression and chemosensitivity by targeting survivin [J].Biochem Biophys Res Commun,2018,501(1): 48-54.
[9] CHAWLA D,KAR R,GUPTA B,et al.Role of survivin and p53 expression in response of primary culture of ovarian cancer cells to treatment with chemotherapeutic agents [J].Int J Gynecol Cancer,2018,28(6):1239-1246.
[10] 侯燕,胡群,熊昊,等.Survivin及其异变体、P53和MDM2在柔红霉素诱导Jurkat细胞凋亡中的变化[J].临床血液学杂志,2013,26(4):482-484,487.
[11] JU R J,CHENG L,QIU X,et al.Hyaluronic acid modified daunorubicin plus honokiol cationic liposomes for the treatment of breast cancer along with the elimination vasculogenic mimicry channels [J].J Drug Target,2018,26(9):793-805.
[12] 陈燕燕,付丽香,周欢,等.EGCG衍生物Y6联合柔红霉素对人肝癌裸鼠移植瘤生长及血管生成的影响[J].广西医科大学学报,2018,35(5):597-602.
[13] HASHEMZAEI M,KARAMI S P,DELARAMIFAR A,et al.Anticancer effects of co-administration of daunorubicin and resveratrol in MOLT-4,U266 B1 and RAJI cell lines[J].Farmacia,2016,64(1):36-42.
[14] 李莉,贾慧,李保平,等.上皮性卵巢癌中Let-7a及其靶基因HMGA2、Cyclin D1的表达[J].郑州大学学报:医学版,2017,52(4):398-402.
[15] WEI M,SHI G,LIU J,et al.Effect of small interfering RNA targeting survivin gene on the expression of survivin and biological function of a human cutaneous squamous cell carcinoma cell line A431[J].Chin J Dermatol,2018,51(4): 305-309.
[16] 杨惠雲,秦国慧,杨黎,等.食管鳞状细胞癌患者外周血和肿瘤组织来源的髓源抑制性细胞中PD-1和TIM-3的表达[J].郑州大学学报:医学版,2019,54(3):336-340.
[17] 徐芝立,张丽红,张翼,等.RNA干扰肾癌Survivin基因表达降低786-O细胞增殖能力[J].河北医科大学学报,2017,38(11):1283-1286,1291.
[18] 刘馨莲,綦黄鹏,魏仙.凋亡抑制因子Survivin的研究进展[J].中外医学研究,2017,15(20):161-163.
[19] 王潇.miR-205通过靶向调控caspase-3表达调节神经胶质瘤的生物学行为[J].河北医科大学学报,2017,38(9):1024-1028,1032.
[20] 张文婷,江莲,朱秀丽,等.地高辛对人B细胞淋巴瘤细胞增殖、凋亡的影响及其分子机制的研究[J].转化医学杂志,2018,7(2):74-78.
[21] KANEKO N,KITA A,YAMANAKA K,et al.Combination of YM155,a survivin suppressant with A STAT3 inhibitor: A new strategy to treat diffuse large B-cell lymphoma[J].Leuk Res,2013,37(9):1156-1161.