敲低长链非编码RNA PURPL-微小RNA-137-Ⅰ型跨膜受体蛋白信号通路抑制乳腺癌的机制  

Knockdown long non-coding RNA PURPL-micro RNA-137-Neurogenic locus notch protein 1 signaling pathway inhibits breast cancer development

作　　者：张松[1] 刘中 姚廷敬[1] 岳喜成[1] Zhang Song;Liu Zhong;Yao Tingjing;Yue Xicheng(Department of Surgical Oncology,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China;Department of General Suegery,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China)

机构地区：[1]蚌埠医学院第一附属医院肿瘤外科,233004 [2]蚌埠医学院第一附属医院普外科,233004

出　　处：《中华实验外科杂志》2022年第10期1895-1898,共4页Chinese Journal of Experimental Surgery

基　　金：蚌埠医学院科技项目(2020byzd134、2020byzd094)。

摘　　要：目的探究敲低长链非编码RNA PURPL调控微小RNA(miR)-137抑制物/Notch同源物1干扰物信号轴对抗乳腺癌的调控机制。方法生信系统分析PURPL与癌症相关性;实时定量反转录聚合酶链反应(RT-qPCR)检测PURPL,miR-137,Ⅰ型跨膜受体蛋白(Notch1)在乳腺癌组织表达;蛋白质印迹法(Western blot)、免疫荧光检测Notch1在乳腺癌组织中蛋白和阳性信号的表达;双荧光素酶报告基因检验miR-137抑制物与PURPL和Notch1的调控机制;细胞计数试剂盒(CCK-8)实验、Transwell实验、细胞伤口愈合实验及原位缺口末端标记法(TUNEL)凋亡分别检测下调PURPL-miR-137-Notch1轴对增殖、侵袭、迁移及凋亡的调控能力;构建乳腺癌裸鼠皮下移植瘤模型检测肿瘤体积和重量,生长情况。两组间比较采用独立样本t检验。结果PURPL和Notch1在乳腺癌组织中高表达(1.01±0.09比2.81±0.22、1.02±0.11比3.19±0.28,t=62.903、59.918,P<0.01),miR-137低表达(0.99±0.08比0.71±0.06,t=23.259,P<0.01);PURPL吸附miR-137,两者竞争性结合,miR-137抑制物逆转siPURPL下调Notch1调控作用(0.47±0.03比0.78±0.06,t=13.864,P<0.01);siPURPL-In-miR-137-siNotch1轴能抑制MCF-7细胞增殖(A值)(2.43±0.37比1.14±0.15,t=9.693,P<0.01)、侵袭细胞数[(75.61±8.78)个比(38.25±3.76)个,t=11.735,P<0.01]、迁移率[(35.56±3.74)%比(20.23±2.61)%,t=10.084,P<0.01]及促进凋亡[(14.91±1.74)%比(19.64±2.33)%,t=4.869,P<0.01];PURPL促进皮下肿瘤体积[(1257.21±135.74)mm^(3)比(485.17±52.62)mm^(3),t=12.990,P<0.01]、重量[(0.28±0.02)g比(0.71±0.08)g,t=12.773,P<0.01],而miR-137则相反,抑制皮下肿瘤体积[(714.38±82.61)mm^(3)比(296.27±27.64)mm^(3),t=11.757,P<0.01]、重量[(0.47±0.05)g比(0.23±0.03)g,t=10.082,P<0.01]低于NC组。结论敲低PURPL可能对抗乳腺癌发生发展有一定的调控作用。Objective To explore the regulatory mechanism of long non-coding RNA PURPL knockdown on microRNA(miR)-137 inhibitor/(Notch homolog 1)Notch1 disruptor signal axis against breast cancer.Methods The relationship between long non-coding RNA PURPL and cancer was explored using bioinformatics;Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of PURPL,miR-137 and Notch1 in breast cancer tissues;Survival curve to observe the correlation between Notch1 and the survival rate of breast cancer patients;Western blot and immunofluorescence were used to detect the expression of Notch1 protein and positive signal in breast cancer tissues;Dual luciferase reporter gene were used to detect the regulatory mechanism of miR-137 inhibitor and PURPL and Notch1;cell counting kit-8(CCK-8)assay,Transwell assay,cell wound healing assay and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)apoptosis were used to detect the regulation of proliferation,invasion,migration and apoptosis of down-regulated PURPL-miR-137-Notch1 axis;The subcutaneous transplanted tumor model of nude mice was constructed to detect tumor volume,weight and growth.Independent sample t test was used for comparison between the two groups.Results PURPL and Notch1 were highly expressed in breast cancer tissues(1.01±0.09 vs.2.81±0.22,1.02±0.11 vs.3.19±0.28,t=62.903,59.918,P<0.01),PURPL adsorb miR-137,and the two competitively bind,miR-137 inhibitor could reverse the regulatory effect of siPURPL on Notch1(0.47±0.03 vs.0.78±0.06,t=13.864,P<0.01);The siPURPL-In-miR-137-siNotch1 axis could inhibit the proliferation(A)(2.43±0.37 vs.1.14±0.15,t=9.693,P<0.01),invasion(cell count)(75.61±8.78 vs.38.25±3.76,t=11.735,P<0.01),migration[(35.56±3.74)%vs.(20.23±2.61)%,t=10.084,P<0.01]and promote apoptosis[(14.91±1.74)%vs.(19.64±2.33)%,t=4.869,P<0.01];PURPL could promote subcutaneous tumor volume[(1257.21±135.74)mm^(3) vs.(485.17±52.62)mm^(3),t=12.990,P<0.01],weight[(0.28±0.02)g vs.(0.71±0.08)g,t=12.773,P<0.01],

关 键 词：乳腺癌 长链非编码RNA PURPL 微小RNA-137 Ⅰ型跨膜受体蛋白 

分 类 号：R737.9[医药卫生—肿瘤]