稳定干扰ITGB3基因可促进人乳腺癌BT549细胞的增殖  

ITGB3 gene-knockdown promotes proliferation of human breast cancer BT549 cells

作　　者：文思阳 徐丽云[1] 杜燕娥[1] 郎磊[1] 孙可馨 阴嘉莉[1] 付立新[1] 席磊[1] 陈燕林[1] 杨丹[1] 柳满然[1] 

机构地区：[1]重庆医科大学教育部临床检验诊断学重点实验室,重庆400016

出　　处：《肿瘤》2016年第5期538-544,共7页Tumor

摘　　要：目的 :探讨稳定干扰整合素β3(integrinβ3,ITGB3)基因对人乳腺癌BT549细胞增殖的影响。方法:采用实时荧光定量PCR法和蛋白质印迹法检测乳腺癌BT549、MCF-7和MDA-MB-453细胞中ITGB3 m RNA和蛋白的表达。将干扰ITGB3表达的LV-ITGB3-sh RNA慢病毒感染BT549细胞后,应用实时荧光定量PCR法和蛋白质印迹法检测BT549细胞中ITGB3 m RNA和蛋白的表达水平,MTT法和FCM法检测BT549细胞的增殖能力和细胞周期,蛋白质印迹法检测BT549细胞中c-Myc和cyclin D1蛋白的表达情况。结果:乳腺癌BT549细胞中ITGB3 m RNA和蛋白的表达水平高于MCF-7和MDA-MB-453细胞(P值均<0.05)。LV-ITGB3-sh RNA慢病毒感染后,BT549细胞中ITGB3 m RNA和蛋白的表达水平低于阴性对照组(LV-NCsh RNA感染BT549细胞)和空白对照组(BT549细胞未进行感染)(P值均<0.01),细胞增殖能力增强(P<0.01),S期细胞所占百分比上升(P<0.01),c-Myc和cyclin D1蛋白的表达水平上调(P值均<0.05)。结论:稳定干扰BT549细胞中ITGB3表达后,可能通过上调c-Myc和cyclin D1蛋白的表达而促进细胞增殖。Objective： To investigate the effect of integrin 133 （ITGB3） gene knockdown on the proliferation of human breast cancer BT549 cells.Methods： The expressions of ITGB3 mRNA and protein in breast cancer BT549, MCF-7 and MDA-MB-453 cells were detected by real- time fluorescent quantitative PCR and Western blotting. After BT549 cells infection with lentivirus containing LV-ITGB3-shRNA with ITGB3 knockdown, the expressions of ITGB3 mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blotting, and the proliferation and cell cycle were measured by MTT and FCM assay, then the expressions of c-Myc and cyclin D1 proteins were determined by Western blotting. Results： The expression levels of ITGB3 mRNA and protein in BT549 cells were higher than those in MCF-7 and MDA-MB-453 cells （all P 〈 0.05）. The expression levels of ITGB3 mRNA and protein in BT549 cells after infection with LV-ITGB3-shRNA lentivirus were lower than those in the negative control （BT549 cells infected with LV-NC-shRNA lentivirus） and the blank control （BT549 cells without any infection） groups （all P 〈 0.01）, the proliferation ability was significantly enhanced （P 〈 0.01）, the percentage of cells in S phase was increased （P 〈 0.01）, and the expressions of c-Myc and cyclin D1 proteins were up-regulated （all P 〈 0.05）. Conclusion： After BT549 cells with stable knockdown can be promoted through up-regulation the express of ITGB3 expression, the cell proliferation ons of c-Myc and cyclin D1 proteins.

关 键 词：乳腺肿瘤 整合素Β3 慢病毒属 RNA干扰 细胞增殖 

分 类 号：R737.9[医药卫生—肿瘤]