microRNA-202激活JNK／SAPK信号通路增强多发性骨髓瘤细胞药物敏感性的研究±  

miR-202 contributes to sensitizing MM cells to drug significantly via activing JNK/SAPK signaling pathway

作　　者：张艳[1] 申娴娟[2] 吴信华[2] 丛辉[1] 倪红兵[1] 鞠少卿[2] 苏建友[1] 

机构地区：[1]南通大学附属医院检验医学中心,226001 [2]南通大学附属医院外科综合实验室,226001

出　　处：《中华血液学杂志》2016年第11期987-992,共6页Chinese Journal of Hematology

基　　金：国家自然科学基金面上项目（81271920、81301498）;江苏省重点研发专项（BE2015654）;江苏省卫计委科研课题（H201422、H201526）

摘　　要：目的研究microRNA-202（miR-202）对多发性骨髓瘤（MM）细胞生长的影响，并初步探讨miR-202在MM细胞药物敏感性中的作用机制。方法荧光定量PCR检测miR-202及其靶基因B淋巴细胞刺激因子（BAFF）在MM细胞中的表达水平。将miR-202模拟物、miR-202抑制物、BAFF干扰质粒（siBAFF）及其阴性对照转染U266细胞，Western blot检测Bcl-2家族和MAPK信号通路蛋白的表达。WST-1法、流式细胞术（Annexin V-FLUOS）分别检测转染后U266细胞的增殖和凋亡情况。结果U266细胞、MM患者CD138＋细胞中miR-202mRNA表达（分别为0．052±0．009、0．304±0．354）均低于健康对照组（3．550±1．126）（P〈0．001，P=0．009），BAFF表达水平（5．700±0．734、9．576±2．887）均高于健康对照组（1．819±0．853）（P〈0．001，P=0．006）。miR-202模拟物转染组细胞增殖抑制率高于对照组[（56．04±0．02）％对（18．89±0．32）％，P=0．002]。Western blot结果显示，转染miR-202模拟物后，U266细胞Bcl-2表达下调约24％，而Bax蛋白的表达上调约1．24倍，miR-202模拟物组细胞凋亡率高于对照组[（49．60±4．89）％对（26．20±1．28）％，P=0．029]。硼替佐米和miR-202模拟物联合组细胞凋亡率为（51．23±5．41）％，高于硼替佐米单独处理组（31．70±4．40）％和硼替佐米与模拟物对照联合处理组[（51．23±5．41）％对（31．70±4．40）％，P=0．047；（51．23±5．41）％对（27．94±4．04）％，P=0．028）]，而miR-202模拟物联合沙利度胺和地塞米松与miR-202模拟物对照组相比差异无统计学意义[（11．66±1．91）％对（10．63±1．74）％，P=0．700；（16．35±1．32）％对（17．43±1．95）％，P=0O．400]。miR-202模拟物联合硼替佐米对U266细胞的增殖抑制率高于硼替佐米单独处理组[（36．93±5．98）％对（18．18±4．10）％，P=0．029]。miR-202模拟物及硼替佐米处理U266细胞后，Objective To explore the role of miR-202 in multiple myeloma （MM） cells, and study the regulation of miR-202 on drug sensitivity of MM cells. Methods miR-202 and BAFF mRNA levels were detected by real-time PCR. U266 cells were transfected with miR- 202- mimics, miR-202- inhibitor, siBAFF and their negative controls. After above treatments, protein levels of Bcl-2 family and MAPK signaling pathway were detected by Western blot analysis, and the proliferation and apoptosis ability of MM cells were examined by WST- 1, Annexin V-FLUOS assay, respectively. Results The results showed that the expression of miR-202 in CD138＋ MM cells （0.304±0.354） and U266 cells （0.052± 0.009） were lower than in normal controls （3.550± 1.126） （P〈0.001, P=0.009）, whereas BAFF mRNA levels （5.700±0.734, 9.576±2.887） were higher than in normal controls （1.819±0.853） （P〈0.001, P= 0.006）. The proliferation ability of U266 cells transfected with miR-202 mimics was significantly inhibited than in control group [ （56.04±0.021）% vs （ 18.89±0.32）%, P=-0.002]. The result of Western blot showed that the expression of Bcl-2 decreased by about 24%, and the expression of Bax increased by about 124% in cells transfected with miR-202 mimics. The apoptosis rate in cells transfected with miR-202 mimics was significantly more than in control group [ （49.60±4.89）% vs （26.20±1.28）%, P=0.029]. The apoptosis rate in miR-202 mimics combined with Bort group （51.23±5.41 ）% was higher as compared with Bort treatment alone （31.70±4.40） % or miR-202 mimics control combined with Bort group （27.94±4.04）%, （P=0.047, P= 0.028）, whereas the apoptosis rate in miR-202 mimics combined with Thai or Dex had no significant difference compared with miR-202 mimics control [ （ 11.66±1.91 ）% vs （ 10.63±1.74）%, P=0.700; （ 16.35± 1.32）% vs （17.43±1.95）%, P=0.400]. The inhibitory rate of cell growth in miR-202 mimics combined with Bort group was higher as compared with Bort treatment

关 键 词：多发性骨髓瘤 微RNAS 抗药性 肿瘤 信号通路 

分 类 号：R733.3[医药卫生—肿瘤]