摘要
目的:探讨mi R-148b是否通过调控Ezrin来调节弥漫性大B细胞淋巴瘤(DLBCL)细胞的药物敏感性。方法:诱导建立人DLBCL耐药细胞株CRL2631/CHOP,实时荧光定量PCR检测CRL2631细胞和CRL2631/CHOP细胞的miR-148b表达,Western blot检测2组细胞的Ezrin蛋白表达;采用双荧光素酶实验检测miR-148b与Ezrin的靶向结合情况;采用miRNA模拟/干扰技术和CCK-8方法检测CRL2631/CHOP细胞和CRL2631细胞对CHOP的敏感性。结果:CRL2631/CHOP细胞mi R-148b表达水平显著低于CRL2631细胞(P=0.001);CRL2631/CHOP细胞Ezrin蛋白表达水平显著高于CRL2631细胞(P<0.05);双荧光素酶实验结果显示,Ezrin是miR-148b的直接靶基因;转染mi R-148b mimic后,CRL2631/CHOP细胞的Ezrin和MDR-1蛋白表达量下降(P<0.05),而CRL2631/CHOP细胞对CHOP敏感性增加(P<0.05);转染miR-148b inhibitor后,CRL2631细胞的Ezrin和MDR-1蛋白表达量上升(P<0.05),而CRL2631细胞对CHOP敏感性下降(P<0.05)。结论:mi R-148b通过调控Ezrin影响DLBCL细胞对CHOP的敏感性。
Objective:To explore the functional role of miR-148 b in regulating chemosensitivity in diffuse large B-cell lymphoma(DLBCL)by targeting Ezrin protein.Methods:After the human DLBCL resistant cell line CRL2631/CHOP was induced,the expression of miR-148 b in CRL2631 cells and CRL2631/CHOP cells was observed by real-time fluorescent quantitative PCR and the difference of Ezrin protein expression between the two groups was measured by Western blot.Double luciferase test was performed to detect the targeted binding between miR-148 b and Ezrin.Sensitivity of CRL2631/CHOP cells and CRL2631 cells to CHOP was determined by microRNAs simulation/interference technique and CCK-8 method.Results:The expression level of miR-148 b in CRL2631/CHOP cells was significantly lower than that in CRL2631 cells(P=0.001).The expression level of Ezrin protein in CRL2631/CHOP cells was obviously higher than that in CRL2631 cells(P<0.05).The results of the double luciferase assay indicated that Ezrin was a direct target gene of miR-148 b.The expression of Ezrin and MDR-1 protein in CRL2631/CHOP cells transfected with miR-148 b mimic were down regulated(P<0.05)and the sensitivity of transfected CRL2631/CHOP cells to CHOP was strengthened(P<0.05).Furthermore,the expression of Ezrin and MDR-1 protein transfected with miR-148 b inhibitor in CRL2631 cells were up-regulated with the sensitivity to CHOP diminished(P<0.05).Conclusion:miR-148 b regulates the chemosensitivity of DLBCL cells to CHOP by targeting Ezrin.
作者
林晓骥
方玮玥
董玉青
孙妮
苏童
杨向绸
郭文坚
何牧卿
姚荣欣
LIN Xiaoji;FANG Weiyue;DONG Yuqing;SUN Ni;SU Tong;YANG Xiangchou;GUO Wenjian;HE Muqing;YAO Rongxin(Department of Haematology,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China)
出处
《温州医科大学学报》
CAS
2020年第1期12-18,共7页
Journal of Wenzhou Medical University
基金
浙江省医药卫生科技计划面上项目（2018KY535）
温州市基础性科研项目（Y20180856）
