转录因子特异性蛋白1对三阴乳腺癌HCC1806细胞增殖凋亡的影响及其机制  

Effect of specificity protein 1 on proliferation,apoptosis andβ-catenin signaling pathway in triple negative breast cancer HCC1806 cells

作　　者：韩亮[1] 孟艳艳 张丽荣[1] 赵峰[3] Han Liang;Meng Yanyan;Zhang Lirong;Zhao Feng(Department of Pathology,China-Japan Union Hospital of Jilin University,Changchun 130033,China;Department of Pharmacy,93011 Unit Hospital of the Chinese People's Liberation Army,Yanji 133000;China Department of Operating Room,China-Japan Union Hospital of Jilin University,Changchun 130033,China)

机构地区：[1]吉林大学中日联谊医院病理科,长春130033 [2]中国人民解放军93011部队医院药房,133000 [3]吉林大学中日联谊医院手术室,长春130033

出　　处：《中华实验外科杂志》2020年第7期1201-1204,共4页Chinese Journal of Experimental Surgery

基　　金：吉林省卫生与健康青年科技骨干培养计划项目(2018Q025)。

摘　　要：目的观察转录因子特异性蛋白1(SP1)对三阴乳腺癌细胞株HCC1806增殖凋亡以及β-连环蛋白(β-catenin)信号通路的影响。方法人正常乳腺上皮细胞株MCF10A和三阴乳腺癌细胞株HCC1806购自中国科学院上海细胞所,通过定量反转录聚合酶链反应(RT-qPCR)和Western blot分析MCF10A和HCC1806细胞中SP1的表达水平,采用Lipofectamine 2000瞬转技术对HCC1806细胞转染SP1小干扰RNA(siRNA)(实验组)和无义siRNA(对照组),通过噻唑蓝(MTT)实验检测SP1对细胞增殖的影响,流式细胞术检测SP1对细胞凋亡的影响,Western blot法检测SP1对细胞β-catenin信号通路相关蛋白[β-catenin、糖原合成酶激酶3β(Gsk3β)、细胞周期蛋白D1(Cyclin D1)、p53和半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3]表达水平的影响。应用SPSS 19.0统计软件进行分析。结果 SP1在三阴乳腺癌细胞株HCC1806中的表达水平(3.09±0.26)明显高于正常乳腺上皮细胞株MCF10A(1.06±0.09,t=0.008,P<0.01)。HCC1806细胞转染SP1 siRNA后,实验组SP1 mRNA相对表达量为(0.37±0.10),SP1 mRNA表达量较对照组下降(66.90±3.79)%,差异有统计学意义(t=0.002,P<0.05)。转染SP1 siRNA后HCC1806细胞实验组增殖率为(40.82±2.67)%,明显低于对照组[(72.72±5.90)%,t=0.000,P<0.05],降低了(43.39±7.65)%,实验组凋亡率为(42.14±4.54)%,显著高于对照组[(22.25±3.07)%,t=0.002,P<0.05],升高了(19.89±5.52)%。实验组β-catenin和Cyclin D1蛋白表达量分别为(0.21±0.08)、(0.70±0.04),对照组β-catenin和Cyclin D1蛋白表达量分别为(0.61±0.09)、(0.92±0.03),实验组β-catenin和Cyclin D1蛋白表达量显著降低(t=0.002、0.001,P<0.05),实验组β-catenin和Cyclin D1蛋白表达量降低分别为(65.80±7.76)%和(23.88±4.99)%,实验组Gsk3β、p53和Caspase-3蛋白表达量分别为(0.73±0.05)、(0.74±0.05)和(0.62±0.11),对照组Gsk3β、p53和Caspase-3蛋白表达量分别为(0.48±0.03)、(0.25±0.03)和(0.36±0.05),实验组Gsk3β、p53和Caspase-3蛋白表达量�Objective:To investigate the effect of specificity protein 1(SP1)on proliferation,apoptosis andβ-catenin signaling pathway in triple negative breast cancer HCC1806 celld.Methods:Human normal breast epithelial cell line MCF10A and triple negative breast cancer cell line HCC1806 were purchased from Shanghai Institute of Cell Science,Chinese Academy of Sciences.The expression of SP1 was detected by real-time quantitative reverse transeriptase-polymerase chain reaction(RT-qPCR)and Western blotting in MCF10A and HCC1806.HCC1806 cells were transfected with SP1 small interfering RNA(siRNA)and scrambled control siRNA by Lipofectamine 2000.The effect of SP1 on cell proliferation and apoptosis was detected by methyl thiazolyl tetrazolium(MTT)assay and flow cytometry.The expression levels ofβ-catenin signaling pathway and downstream related proteins[β-catenin,glycogen synthase kinase 3β(Gsk3β),Cyclin D1,p53 and cysteinyl aspartate-specific protease(Caspase)-3]in the HCC1806 cells were detected by Western blotting.SPSS 19.0 statistical software was used for analysis.Results:The expression of SP1 in HCC1806 cells(3.09±0.26)was significantly higher than that in MCF10A(1.06±0.09,t=0.008,P<0.01).The relative expression of SP1 mRNA of HCC1806 cells transfected with SP1 siRNA in the experimental group was(0.37±0.10),SP1 expression of HCC1806 cells transfected with SP1 siRNA significantly decreased(t=0.002,P<0.05),by(66.90±3.79)%compared with the control group.After transfection with SP1 siRNA,the proliferation rate of the experimental group was(40.82±2.67)%,significantly lower than that of the control group[(72.72±5.90)%,t=0.000,P<0.05],and the apoptosis rate of the experimental group was(42.14±4.54)%,significantly higher than that of the control group[(22.25±3.07)%,t=0.002,P<0.05],and increased(19.89±5.52)%.The protein expression levels ofβ-catenin and Cyclin D1 in the experimental group were(0.21±0.08)and(0.70±0.04),respectively;the protein expression levels ofβ-catenin and Cyclin D1 in the control group were(0

关 键 词：乳腺癌 转录因子特异性蛋白1 增殖 脱噬作用 β-连环蛋白信号通路 

分 类 号：R737.9[医药卫生—肿瘤]