lncRNA MEG3调控NLRP3/caspase-1/GSDMD通路影响三阴性乳腺癌对紫杉醇的敏感性  

lncRNA MEG3 regulates the NLRP3/caspase-1/GSDMD pathway to affect the sensitivity of triple-negative breast cancer to paclitaxel

作　　者：于冰 杨旭 陈之梦 雷萌[2] YU Bing;YANG Xu;CHEN Zhimeng;LEI Meng(Center for Drug Evaluation,National Medical Products Administration,Beijing 100022,China;College of Science,Nanjing Forestry U-niversity,Jiangsu Nanjing 210037,China)

机构地区：[1]国家药品监督管理局药品审评中心,北京100022 [2]南京林业大学理学院,江苏南京210037

出　　处：《现代肿瘤医学》2023年第4期597-602,共6页Journal of Modern Oncology

基　　金：国家自然科学基金面上项目(编号:21877061)。

摘　　要：目的:探究长链非编码RNA(long non-coding RNA,lncRNA)母系表达基因3(maternally expressed gene 3,MEG3)调控含NLR家族Pyrin域蛋白3(NLR family,Pyrin domain containing protein 3,NLRP3)/含半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase 1,caspase-1)/消皮素D(gasdermin D,GSDMD)通路对三阴性乳腺癌(triple-negative breast cancer,TNBC)对紫杉醇(paclitaxel,PTX)敏感性的影响。方法:通过逐渐增加PTX剂量间歇作用的方法诱导TNBC耐药细胞MDA-MB-231/R,qRT-PCR检测lncRNA MEG3表达;将MDA-MB-231细胞分为对照组(未转染+PTX)、Vector组(空载体+PTX)、pcDNA3.1-MEG3组(pcDNA3.1-MEG3表达载体+PTX)、pcDNA3.1-MEG3+BAY11-7082组(pcDNA3.1-MEG3表达载体+PTX+5μmol/L NLRP3抑制剂BAY11-7082),qRT-PCR检测转染后lncRNA MEG3表达;CCK-8法检测MDA-MB-231细胞增殖情况;通过免疫荧光染色和扫描电镜(scanning electron microscope,SEM)观察MDA-MB-231细胞焦亡情况;Western blot检测MDA-MB-231细胞中NLRP3/caspase-1/GSDMD通路蛋白表达;体内成瘤实验检测肿瘤质量。结果:与MDA-MB-231细胞相比,MDA-MB-231/R细胞中lncRNA MEG3表达水平显著降低(P<0.05);与对照组相比,pcDNA3.1-MEG3组细胞增殖抑制率、GSDMD-N+细胞数量、细胞焦亡、细胞凋亡率及NLRP3、cleaved-caspase 1/caspase-1、GSDMD-N/GSDMD表达水平显著增加,IC50、肿瘤质量显著降低(P<0.05);与pcDNA3.1-MEG3组相比,pcDNA3.1-MEG3+BAY11-7082组细胞增殖抑制率、GSDMD-N+细胞数量、细胞焦亡、细胞凋亡率及NLRP3、cleaved-caspase 1/caspase-1、GSDMD-N/GSDMD表达水平显著降低,IC50、肿瘤质量显著增加(P<0.05)。结论:上调lncRNA MEG3表达可通过激活NLRP3/caspase-1/GSDMD通路,促进MDA-MB-231细胞焦亡,以此增加TNBC对PTX的敏感性。Objective:To explore the effect of long non-coding RNA(lncRNA)maternally expressed gene 3(MEG3)on the sensitivity of triple-negative breast cancer(TNBC)to paclitaxel(PTX)by regulating the NLR family,Pyrin domain containing protein 3(NLRP3)/cysteinyl aspartate specific proteinase 1(caspase-1)/gasder⁃min D(GSDMD)pathway.Methods:TNBC drug-resistant cells MDA-MB-231/R were induced by gradually in⁃creasing the dose of PTX intermittent action method,and qRT-PCR was used to detect the expression of lncRNA MEG3.MDA-MB-231 cells were divided into control group(untransfected+PTX),Vector group(empty vector+PTX),pcDNA3.1-MEG3 group(pcDNA3.1-MEG3 expression vector+PTX),pcDNA3.1-MEG3+BAY11-7082 group(pcDNA3.1-MEG3 expression vector+PTX+5μmol/L NLRP3 inhibitor BAY11-7082).qRT-PCR was used to detect the expression of lncRNA MEG3 after transfection.CCK-8 method was used to detect the prolifer⁃ation of MDA-MB-231 cells.The pyroptosis of MDA-MB-231 cells was observed by immunofluorescence stai⁃ning and scanning electron microscope(SEM).Western blot was used to detect the protein expression of NLRP3/caspase-1/GSDMD pathway in MDA-MB-231 cells.In vivo tumorigenic assay was used to detect tumor mass.Re⁃sults:Compared with MDA-MB-231 cells,the expression level of lncRNA MEG3 in MDA-MB-231/R cells was significantly reduced(P<0.05).Compared with the control group,the cell proliferation inhibition rate,number of GSDMD-N+cells,cell pyroptosis,cell apoptosis rate and the expression level of NLRP3,cleaved-caspase 1/caspase-1,GSDMD-N/GSDMD proteins in the pcDNA3.1-MEG3 group increased significantly,and IC50,tumor mass re⁃duced significantly(P<0.05).Compared with the pcDNA3.1-MEG3 group,the cell proliferation inhibition rate,number of GSDMD-N+cells,cell pyroptosis,cell apoptosis rate and the expression level of NLRP3,cleaved-caspase 1/caspase-1,GSDMD-N/GSDMD proteins in the pcDNA3.1-MEG3+BAY11-7082 group reduced significantly,and IC50,tumor mass increased significantly(P<0.05).Conclusion:Up-regulating the expression of lncRNA MEG3

关 键 词：母系表达基因3 含NLR家族Pyrin域蛋白3/含半胱氨酸的天冬氨酸蛋白水解酶/消皮素D 三阴性乳腺癌 紫杉醇 敏感性 

分 类 号：R737.9[医药卫生—肿瘤]