靶向EGFR的miR-7-5p对人乳腺癌细胞增殖和侵袭的影响  

Effect of miR-7-5p targeting EGFR on proliferation and invasion of human breast cancer cell lines

作　　者：林维佳[1] 郑琳[1] 代建波[1] 许澍阳 

机构地区：[1]常州市第二人民医院乳腺外科,江苏常州213000

出　　处：《中华肿瘤防治杂志》2015年第15期1176-1179,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的验证靶向表皮生长因子(epidermal growth factor receptor,EGFR)的微RNA-7-5p(microRNA-7-5p,miR-7-5p)对人乳腺癌细胞MDA-MB-231的增殖和侵袭能力的影响。方法应用双荧光素酶检测试剂验证EGFR为miR-7-5p的下游靶基因;在人乳腺癌细胞MDA-MB-231中转染miR-7-5p阴性对照质粒和pMIR-report-3′UTR/3′UTRm质粒,观察细胞中EGFR的mRNA水平和蛋白水平的表达变化;通过CCK-8实验,观察转染pMIR-report-3′UTR/3′UTRm质粒后细胞增殖能力的变化;利用流式细胞术检测转染pMIR-report-3′UTR/3′UTRm质粒后MDA-MB-231细胞周期的改变。结果与对照组相比,miR-7-5p可显著降低非突变载体的相对荧光素酶活性,χ2=32.6,P<0.01。miR-7-5p靶向EGFR可使其mRNA表达水平比空白对照下调81.2%,χ2=136.1,P<0.05;比阴性对照下调79.1%,χ2=130.6,P<0.05。其蛋白水平则比空白对照下调71.0%,χ2=110.1,P<0.05;比阴性对照下调70.0%,χ2=107.7,P<0.05。过表达miR-7-5p第3天后,MDA-MB-231细胞的增殖能力比空白对照组降低29.3%,χ2=33.9,P<0.05;比阴性对照组降低27.5%,χ2=32.2,P<0.05。而细胞周期则表现为G0/G1期阻滞。相比空白对照组,实验组细胞的转移能力降低了约48.9%,χ2=64.9,P<0.05;相比阴性对照组则降低了50.9%,χ2=68.5,P<0.05。侵袭能力相比空白对照组降低了约45.7%,χ2=59.8,P<0.05;相比阴性对照组则降低了43.7%,χ2=56.4,P<0.05。结论靶向EGFR的miR-7-5p可抑制人乳腺癌细胞的增殖能力和侵袭能力。OBJECTIVE To investigate the effect of microRNA-7-Sp （miR-7-5p） targeting epidermal growth factor receptor （EGFR） on proliferation and invasion of human breast cancer cell lines MDA-MB-231. METHODS The interac- tion between EGFR and miR-7-Sp was validated through the dual lueiferase reporter assay. The effects of miR-7-Sp on EGFR and on proliferation were measured by immunoblot, CCK-8 assay, flow cytometry under transfection of miR-7-Sp/ N.C. and pMIR-report-3＂UTR/3＂UTRm plasmid in MDA-MB-231 breast cancer cell lines. RESULTS MiR-7-Sp targeted EGFR expression was significantly decreased, comparing to control group （χ2=32.6, P〈0.01）. As compared to control group and negative group, up-regulate miR-7-Sp level in breast cancer cell line MDA-MB-231 resulted in attenuated mR- NA expression 81. 2% （Xz=136.1,P〈0.05） and 79.1% （χ2=130.6,P〈0.05）, and attenuated protein expression 71.0% （χ2 =110.1,P〈0.05） and 70.0% （χ2 = 107.7, P〈0.05） as well as decreased proliferation capacity 29.3% （χ2 =33.9,P〈0.05） and 27.5% （χ2 =32.2,P〈0.05）. Additionally, compared to control group and negative group, the migration capacity of MDA-MB-231 cell was decreased in 48.9% （χ2 =64.9,P〈0.05） and 50.9% （χ2 =68.5, P〈 0.05）, and the invasion capacity was decreased in 45.7% （χ2=59.8, P〈0.05） and 43.7% （χ2=56.4, P〈0.05）, which was resulted from transfecting with miR-7-5p. CONCLUSION The miR-7-5p targeting EGFR can suppress prolif- eration and invasion of human breast cancer cell line MDA-MB-231.

关 键 词：miR-7-5p EGFR 乳腺癌 细胞增殖 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]