弥漫性大B细胞淋巴瘤治疗中利妥昔单抗耐药与Smarcal1、PP2A-B55-α、PP2A-B56-α蛋白表达的相关性  

Correlation of rituximab resistance with the expressions of Smarcal 1,PP2A-B55-αand PP2A-B56-αin the treatment of diffuse large B-cell lymphoma

作　　者：刘静 邓槿 顾季炜 马欣玥 宋国齐 LIU Jing;DENG Jin;GU Jiwei;MA Xinyue;SONG Guoqi(Medical College of Nantong University,Nantong 226001,Jiangsu;Department of Clinical Laboratory,Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu,China;Department of Hematology,Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu,China)

机构地区：[1]南通大学医学院,江苏南通226001 [2]南通大学附属医院检验科,江苏南通226001 [3]南通大学附属医院血液科,江苏南通226001

出　　处：《临床检验杂志》2021年第6期418-423,共6页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金(8177011136)。

摘　　要：目的探讨弥漫大B细胞淋巴瘤(DLBCL)治疗中利妥昔单抗耐药与Smarcal1、PP2A-B55-α、PP2A-B56-α蛋白表达的相关性。方法选取2种“双重打击”淋巴瘤衍生的细胞系,活化B细胞样(ABC型)OCI-LY10(LY10)和生发中心B细胞样型(GCB型)OCI-LY18(LY18),利用“梯度加药法”构建利妥昔单抗耐药细胞系(LY10RR及LY18RR);采用CCK-8法检测不同浓度利妥昔单抗分别干扰亲本及耐药细胞后的细胞存活率并计算半数生长抑制浓度值(IC_(50));AnnexinⅤ/PI双染法检测50μg/mL利妥昔单抗分别干扰亲本及耐药细胞48 h后的凋亡比率;western blot检测细胞耐药后对Smarcal1、PP2A-B55-α和PP2A-B56-α蛋白表达的影响;在耐药细胞系中分别干扰Smarcal1和PP2A-B55-α的表达以及过表达PP2A-B56-α,采用CCK-8法检测不同浓度利妥昔单抗对转染后耐药细胞活力的影响。结果经不同浓度利妥昔单抗处理48 h后,LY10及LY10RR细胞IC_(50)分别为(0.91±0.07)μg/mL和(287.40±26.30)μg/mL,耐药指数为315.58;LY18及LY18RR的IC_(50)分别为(4.73±0.24)μg/mL和(532.10±37.95)μg/mL,耐药指数为112.41,二者差异具有统计学意义(t值分别为10.51和15.18,P均<0.001);经50μg/mL利妥昔单抗处理48 h后,LY10RR和LY18RR的凋亡率与对照组的细胞凋亡率相比,差异均无统计学意义(t值分别为2.86和2.90,P值分别为0.06和0.05),而利妥昔单抗处理48 h后,LY10和LY18的凋亡率明显高于对照组细胞的凋亡率,且差异有统计学意义(t值分别为10.50和9.86,P均<0.001)。与亲本细胞相比,在发生利妥昔单抗耐药细胞系中,Smarcal1和PP2A-B55-α表达量增加,PP2A-B56-α表达水平下降,且差异均具有统计学意义(P均<0.05)。耐药细胞系转染并经不同浓度利妥昔单抗处理后,sh-Smarcal1组、sh-PP2A-B55-α组细胞存活率均明显低于sh-NC组,oe-PP2A-B56-α组细胞存活率明显低于oe-vector组(P均<0.01)。结论成功构建并鉴定稳定的对利妥昔单抗耐药的LY10RR和LY18RR细胞模型,并�Objective To investigate the correlation of rituximab resistance with the expressions of Smarcal1,PP2A-B55-αand PP2A-B56-αin the treatment of diffuse large B-cell lymphoma(DLBCL).Methods Two cell lines derived from"double strike"lymphoma,including activated B-cell-like(ABC)OCI-LY10(LY10)and germinal center B-cell-like(GCB)OCI-LY18(LY18),were selected to construct the rituximab-resistant DLBCL cell lines(LY10RR and LY18RR)by the concentration gradient dosing method.The survival rates of parental and drug-resistant cells interfered with different concentrations of rituximab were detected by the CCK-8 method,and the half maximal inhibitory concentration(IC_(50))was calculated.The apoptotic rate of parental and drug-resistant cells interfered by 50μg/mL of rituximab for 48 hours were determined by the AnnexinⅤ/PI double staining method.The effects of cell resistance on the expressions of Smarcal 1,PP2A-B55-αand PP2A-B56-αwere deteced by western blot.The effects of different concentrations of rituximab on the viability of drug-resistant cells transfected with sh-Smarcal 1,sh-PP2A-B55-αand oe-PP2A-B56-αwere detected by the CCK-8 method.Results The IC_(50)of LY10 and LY10RR cells treated with different concentrations of rituximab for 48 hours were(0.91±0.07)μg/mL and(287.40±26.30)μg/mL,respectively,and the drug resistance index was 315.58.The IC_(50)of LY18 and LY18RR cells were(4.73±0.24)μg/mL and(532.10±37.95)μg/mL,respectively,and the drug resistance index was 112.41.There were significant differences between LY10 and LY10RR and between LY18 and LY18RR(t=10.51 and 15.18,respectively,P<0.001).The apoptosis rates of LY10RR and LY18RR cells treated with 50μg/mL of rituximab for 48 hours showed no statistical significance with that of the control group(t=2.86 and 2.90,respectively;P=0.06 and 0.05,respectively).However,the apoptosis rates of LY10 and LY18 cells treated with 50μg/mL of rituximab for 48 hours were significantly higher than that of the control group(t=10.50 and 9.86,respectively,P<0.001).C

关 键 词：弥漫大B细胞淋巴瘤 利妥昔单抗 MYC Smarcal1 PP2A 

分 类 号：R446[医药卫生—诊断学] R55[医药卫生—临床医学]