Notch1调控PTEN表达对非小细胞肺癌细胞干性及阿霉素耐药性的影响  

Effects of Notch1 regulating PTEN expression on stemness of non-small cell lung cancer cells and adriamycin resistance

作　　者：张媛媛 马静 何会娜[3] 郭银[4] 李香兰[1] 池菲[1] ZHANG Yuanyuan;MA Jing;HE Huina;GUO Yin;LI Xianglan;CHI Fei(Department of Emergency,Hebei Chest Hospital,Shijiazhuang 050041,China;Department of Gastroenterology,Hebei Chest Hospital;Department of Nursing,Hebei Chest Hospital;Department of Radiotherapy,Fourth Hospital of Hebei Medical University)

机构地区：[1]河北省胸科医院急诊科,石家庄050041 [2]河北省胸科医院消化内科 [3]河北省胸科医院护理部 [4]河北医科大学第四医院放疗科

出　　处：《山西医科大学学报》2021年第10期1247-1253,共7页Journal of Shanxi Medical University

基　　金：河北省医学科学研究课题计划项目(20191015,20191011)。

摘　　要：目的对Notch1进行表达沉默后,探究其是否可能通过调控人第10号染色体缺失的磷酸酶(PTEN)表达对非小细胞肺癌A549细胞干性及阿霉素耐药性产生影响。方法将A549细胞设置为对照组、NC-shRNA组(转染NC-shRNA)、Notch1-shRNA组(转染Notch1-shRNA)、Notch1-shRNA+bpV(phen)组(转染Notch1-shRNA后添加100 nmol/L PTEN抑制剂bpV(phen)),采用qRT-PCR检测A549细胞中Notch1 mRNA表达情况。肿瘤成球实验检测肿瘤成球情况。细胞划痕实验检测A549细胞迁移情况。Western blot检测A549细胞内八聚体结合转录因子4(OCT4)、CD133、醛脱氢酶1A1(ALDH1A1)、PTEN、c-Myc、细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶2(MMP-2)蛋白表达情况。MTT检测A549细胞阿霉素耐药性。结果与对照组相比,Notch1-shRNA组肿瘤成球率、细胞迁移率、存活率、MMP-9、MMP-2、OCT4、CD133、ALDH1A1、c-Myc、CyclinD1表达水平、阿霉素耐药性均降低(P<0.05),PTEN表达水平增加(P<0.05)。与Notch1-shRNA组相比,Notch1-shRNA+bpV(phen)组肿瘤成球率、细胞迁移率、存活率、MMP-9、MMP-2、OCT4、CD133、ALDH1A1、c-Myc、CyclinD1表达水平、阿霉素耐药性均显著增加,而PTEN表达水平显著降低(P<0.05)。结论Notch1表达下调可通过促进PTEN表达抑制A549细胞干性及阿霉素耐药性。Objective To explore the effect of Notch1 silencing on the stemness and adriamycin resistance of non-small cell lung can-cer A549 cells by regulating the expression of gene of phosphate and tension homology deleted on chromsome ten(PTEN).Methods A549 cells were divided into control group,NC-shRNA group,Notch1-shRNA group,Notch1-shRNA+bpV(phen)group.A549 cells were given 100 nmol/L PTEN inhibitor bpV(phen)after transfection of Notch1-shRNA in Notch1-shRNA+bpV(phen)group.The expression of Notch1 mRNA in A549 cells was detected by qRT-PCR.Tumor sphere formation was detected by tumor sphere formation assay.The migration of A549 cells was detected by cell scratch test.The protein expression of octamer-binding transcription factor(OCT4),CD133,aldehyde dehydrogenase 1A1(ALDH1A1),PTEN,c-Myc,CyclinD1,matrix metallopeptidase 9(MMP-9),matrix metallopeptidase 2(MMP-2)in A549 cells was detected by Western blot.The drug resistance of A549 cells to adriamycin was detected by MTT.Results Compared with control group,the tumor sphere formation rate,the cell migration rate,the survival rate,the expression levels of MMP-9,MMP-2,OCT4,CD133,ALDH1A1,c-Myc,CyclinD1,and the adriamycin resistance were decreased in Notch1-shRNA group(P<0.05),while the PTEN expression level was increased(P<0.05).Compared with Notch1-shRNA group,the tumor sphere formation rate,the cell migration rate,the survival rate,the expression levels of MMP-9,MMP-2,OCT4,CD133,ALDH1A1,c-Myc,CyclinD1,and adriamycin resistance were significantly increased in Notch1-shRNA+bpV(phen)group(P<0.05),while the PTEN expression level was significantly decreased(P<0.05).Conclusion Down-regulation of Notch1 expression can inhibit the stemness of A549 cells and adriamycin resistance by promoting PTEN expression.

关 键 词：非小细胞肺癌 A549细胞 细胞干性 阿霉素 耐药性 

分 类 号：R734.2[医药卫生—肿瘤]