转染微小RNA125b抑制序列的三阴性乳腺癌细胞增殖和凋亡变化  

Effect of miR-125b on proliferation and apoptosis of human triple negative breast cancer cells MDA-MB-468

作　　者：聂俊[1] 江波[1] 何文杰[1] 朱颖 涂长玲[1] 董坚[1] NIE Jun;JIANG Bo;HE Wenfie;ZHU Ying;TU Changling;DONG Jian(Yunnan Cancer Hospital, Kunming 650118, China)

机构地区：[1]云南省肿瘤医院,昆明650118

出　　处：《山东医药》2018年第19期18-21,共4页Shandong Medical Journal

基　　金：云南省科技厅-昆明医科大学应用基础研究联合专项资金项目(U0120160174)

摘　　要：目的观察转染微小RNA125b(miR-125b)抑制序列的三阴性乳腺癌(TNBC)细胞MDA-MB-468增殖及凋亡情况的变化。方法将MDA-MB-468细胞分为miR-125b抑制序列转染组、无关序列转染组和空白对照组,miR-125b抑制物转染组、无关序列转染组分别转染miR-125b抑制序列和无关序列,空白对照组不转染。转染48 h后采用实时荧光定量PCR法检测3组细胞中的miR-125b;转染后24、48、72、96、120 h分别采用MTT法观察各组细胞增殖情况(以OD570表示);转染72 h后采用流式细胞术测算各组细胞凋亡率;各组细胞长至80%~90%汇合时采用Western blotting法检测NF-κBp65和Caspase-3蛋白。结果转染96、120 h时miR-125b抑制序列转染组细胞OD570低于无关序列转染组和空白对照组(P均<0.05)。转染72 h后miR-125b抑制序列转染组、无关序列转染组和空白对照组细胞凋亡率分别为24.85%±2.21%、1.47%±0.78%和1.32%±0.65%。miR-125b抑制序列转染组细胞凋亡率与无关序列转染组和空白对照组相比,P均<0.05,无关序列转染组和空白对照组细胞凋亡率相比,P>0.05。与无关序列转染组和空白对照组相比,miR-125b抑制序列转染组细胞NF-κBp65相对表达量下降(P均<0.05),Caspase-3相对表达量上升(P均<0.05)。结论转染miR-125b抑制序列的TNBC细胞MDA-MB-468增殖受到抑制、凋亡增强。这可能与转染miR-125b抑制序列下调MDA-MB-468细胞NF-κBp65和上调Caspase-3蛋白表达有关。Objective To observe the proliferation and apoptosis of triple negative breast cancer（ TNBC） cells MDAMB-468 transfected with microRNA125 b（ miR-125 b） inhibitor. Methods MDA-MB-468 cells were divided into the miR-125 b inhibitor group,negative control group,and blank control group. MDA-MB-468 cells of the miR-125 b inhibitor group and negative control group were transfected with miR-125 b inhibitors and nonsense sequence,respectively. At 48 h after transfection,miR-125 b in the three groups was detected by real-time fluorescent quantitative PCR to evaluate transfection efficiency. At 24,48,72,96,and 120 h after transfection,MTT was utilized to observe the proliferation（ expressed by OD570）. The apoptosis rate was detected by flow cytometry. Western blotting was used to determine the protein expression of Caspase-3 and NF-κBp65. Results MTT assay showed that inhibiting miR-125 b expression suppressed the proliferation of MDA-MB-468 cells at 96 and 120 h after transfection,the OD570 of miR-125 b inhibitor group were 0. 32 ± 0. 01 and 0. 37 ± 0. 01,which were lower than that of the negative control group（ 0. 58 ± 0. 02 and 0. 74 ± 0. 01） and the blank control group（ 0. 61 ± 0. 01 and 0. 75 ± 0. 02）（ both P〈0. 05）. At 72 h after transfection,the apoptosis rate of the miR-125 b inhibitor group,negative control group,and blank control group were 24. 85% ± 2. 21%,1. 47% ± 0. 78%,and1. 32% ± 0. 65%,respectively. Significant difference was found between the miR-125 b inhibitor group,negative control group,and blank control group（ P〈0. 05）; no significant difference was found between the negative control group and blank control group（ P〈0. 05）. Western blotting showed that the protein expression of NF-κBp65 were down-regulated（ 0. 48 ± 0. 02） and the Caspase-3 was up-regulated（ 1. 02 ± 0. 03） in the miR-125 b inhibitor group as compared with that of the negative control group and blank control group（ both P〈0. 05）. Conclusion Inhibiting miR-125 b expression

关 键 词：微小RNA 微小RNA125b 三阴性乳腺癌 细胞增殖 细胞凋亡 核因子-ΚB 含半胱氨酸的天冬氨酸蛋白水解酶3 

分 类 号：R737.9[医药卫生—肿瘤]