Linc00339对三阴乳腺癌MDA-MB-231细胞生物学特性的影响  

Lind00339 promotes the development and progression of triple-negative breast cancer via miR-135b/APC-mediated Wnt/p-catenin signaling pathway

作　　者：许迪锋 罗璟璐 章恒[3] Xu Difeng;Luo Jinglu;Zhang Heng(Department of General Surgery,Dajiangdong Hospital,Hangzhou 311225,China;Department of Opera tire Anesthesiology,Reproductive Health Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830000,China;Radiotherapy Department,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China)

机构地区：[1]杭州市大江东医院普外科,311225 [2]新疆维吾尔自治区生殖健康医院手术麻醉科,乌鲁木齐830000 [3]新疆维吾尔自治区人民医院放疗一科,乌鲁木齐830001

出　　处：《中国医师杂志》2020年第10期1505-1510,1515,共7页Journal of Chinese Physician

基　　金：新疆维吾尔自治区自然科学基金项目(2019D01C258)。

摘　　要：目的探讨Linc00339对三阴乳腺癌发生发展的作用及其相关分子机制。方法采用实时荧光定量PCR(qRT-PCR)法检测人乳腺正常上皮细胞和乳腺癌细胞中Linc00339的表达水平;采用质粒转染试验在MDA-MB-231细胞中过表达Linc00339;采用四甲基偶氮唑盐(MTT)法检测MDA-MB-231细胞的活性;采用黏附试验、Transwell试验分别检测MDA-MB-231细胞的黏附、侵袭和迁移能力;采用Western blot检测MDA-MB-231细胞中APC、Wnt/β-catenin信号通路相关蛋白的表达水平;采用qRT-PCR法检测MDA-MB-231细胞中miRNA-135a、miRNA-135b及miRNA-138的表达水平;将体外转染成功的MDA-MB-231细胞接种于裸鼠体内建立荷瘤裸鼠模型,观察和测量肿瘤块的体积和重量,采用Western blot检测裸鼠瘤体组织中APC、Wnt/β-catenin信号通路相关蛋白的表达水平,采用qRT-PCR法检测裸鼠瘤体组织中miRNA-135a、miRNA-135b及miRNA-138的表达水平。结果与乳腺正常上皮细胞组(Hs578Bst)相比,乳腺癌细胞组(MCF-7、MDA-MB-468、MDA-MB-231)的Linc00339表达水平均显著上调,以三阴乳腺癌MDA-MB-231细胞尤为显著;转染实验结果表明与pcDNA3.1组相比,Linc00339转染显著上调Linc00339表达水平并可显著增加MDA-MB-231细胞的活力;Linc00339过表达能够显著增加MDA-MB-231细胞的增殖、黏附、迁移和侵袭能力,增加荷瘤裸鼠肿瘤块的体积和重量;Western blot结果均表明体内外Linc00339过表达能够下调APC蛋白的表达和上调Wnt/β-catenin信号蛋白的表达,同时qRT-PCR结果表明Linc00339过表达能够上调miRNA-135b表达水平,而对miRNA-135a和miRNA-138无影响。结论 Linc00339可能通过miR-135b/APC介导的Wnt/β-catenin信号通路促进三阴乳腺癌的发生和发展。Objective To investigate the role of LincOO339 in the development of triple-negative breast cancer and its related molecular mechanisms.Methods The expression levels of Linc00339 in human nonnal breast epithelial cells and breast cancer cells were detected by real time fluorescent quantitative polymerase chain reaction(qRT-PCR).The LincOO339 was overexpressed in MDA-MB-231 cells by plasmid transfection test;the activity of MDA-MB-231 cells was detected by methyl thiazolyl tetrazolium(MTT)method;the adhesion,invasion and migration ability of MDA-MB-231 cells were detected by adhesion test and transwell test respectively;Western blot was used to detect the expression of APC and Wnt/p-catenin signaling pathway-related proteins in MDA-MB-231 cells.qRT-PCR was used to detect the expression of miRNA-135a,miRNA-135b and miRNA-138 in MDA-MB-231 cells.MDA-MB-231 cells were successfully transfected into nude mice to establish tumor-bearing nude mice model.The volume and weight of tumor were observed and measured.The expression levels of A PC,Wnt/0-catenin signali ng pathway related proteins were detected by Western blot,and the expression levels of miRNA-135a,miRNA-135b and miRNA-138 were detected by qRT-PCR.Results Compared with the normal breast epithelial cell group(Hs578Bst),the expression level of Linc00339 in breast c ancer cell lines(MCF-7,MDA-MB-468,MDAMB-231)was significantly up-regulated,especially MDA-MB-231.The results of transfection experiments showed that the expression level of LincOO339 and cell viability were significantly up-regulated in the LincOO339 group compared with the pcDNA3.1 group.The overexpression of LincOO339 significantly increased the proliferation,adhesion and migration and invasion ability of MDA-MB-231 cells,as well as increased the volume and weight of tumor mass in tumor-bearing nude mice.Western blot results showed that Linc00339 ovrrexpression can down-regulate A PC protein expression and up-regulate Wnt/β-catenin signaling protein expression.Mean while,qRT-PCR results indicated Lin

关 键 词：三阴性乳腺癌 细胞系 肿瘤 RN A 长链非编码 Wnt/p-eatenin信号通路 

分 类 号：R737.9[医药卫生—肿瘤]