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机构地区:[1]湖北医药学院附属太和医院普外3科,湖北十堰442000 [2]湖北医药学院基础医学研究所,湖北十堰442000
出 处:《现代肿瘤医学》2012年第12期2476-2479,共4页Journal of Modern Oncology
摘 要:目的:探讨利用RNA干扰沉默Delta样配体4(Delta-like ligand 4,DLL4)对MCF-7细胞生长、凋亡的影响,为乳腺癌的基因治疗提供一个可选择的靶点和方法。方法:针对DLL4基因的DNA设计具有特异性的SiRNA,经脂质体转染MCF-7,采用免疫组化方法检测空脂质体组和转染组DLL4基因表达有否阻断。用MTT法测定MCF-7细胞生长曲线,流式细胞仪检测细胞凋亡率及细胞周期改变。结果:构建DLL4-SiR-NA表达载体能够有效抑制MCF-7中DLL4的表达(P<0.05)。RNA干扰沉默DLL4基因可抑制MCF-7细胞增殖,促进细胞凋亡,G2/M期细胞比例明显增加(P<0.05)。结论:RNAi技术能够有效抑制DLL4基因的表达,抑制DLL4基因的表达可抑制MCF-7细胞增殖,促进MCF-7细胞凋亡。Objective:To inhibit the expression of DLL4 gene in human breast carcinoma cell MCF-7 cells by SiRNA on proliferation and apoptosis.Methods: The specific SiRNA was designed for DLL4 gene and transfected into human breast carcinoma cell MCF-7 cells with liposomal,to study whether the expression of DLL4 gene can be blocked.Immunohistochemical method was used to detect in contol and transfected groups.Proliferation of MCF-7 cells was examined by MTT assay.Apoptosis of MCF-7 cells was examined by FCM.Cell cycle distribution of MCF-7 cells was analyzed by FCM.Results: The constructed vector could effectively inhibit the expression of DLL4 in MCF-7 cells.Knockdown of Notch ligand DLL4 expression by RNAi could reduce proliferation and induce apoptosis in the MCF-7 cells.Decreased DLL4 expression could lead to cell cycle G2/M arrest.Conclusion: The RNA interference technology could effectively block the DLL4 gene expression.Inhibition of Dll4/Notch signaling pathway can lead to inhibition of cancer cell proliferation and a concomitant increase in cells undergoing apoptosis,and thus,Dll4 would be an important target for cancer therapeutic approach.
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