腺病毒介导靶向NF-κB基因的shRNA对体外食蟹猴子宫内膜细胞增殖活性的影响  

Adenovirus vector-mediated short hairpin RNA targeting nuclear factor-κB suppresses proliferation of endometrial cells of Macaca fascicularis in vitro

作　　者：朱峰城 刘木彪[1] 潘颖[1] 王雪峰[1] 陈燕英[1] 

机构地区：[1]南方医科大学珠江医院妇产科,广东广州510282

出　　处：《南方医科大学学报》2015年第3期390-396,共7页Journal of Southern Medical University

基　　金：广州市科技计划重大民生攻关专项(11Bpp ZXaa2120016)

摘　　要：目的合成高特异性靶向NF-κB基因的sh RNA腺病毒载体,观察其对体外培养的食蟹猴子宫内膜细胞的增殖活动是否具有直接的抑制效应。方法设计携带有NF-κB-p65基因及空载基因的sh RNA腺病毒载体,体外培养食蟹猴子宫内膜细胞,分成实验组及对照组,实验组转染携带NF-κB-p65 sh RNA的腺病毒,对照组转染携带空载基因的腺病毒,共培养后观察有关细胞增殖活性如凋亡蛋白及细胞周期的改变情况。结果腺病毒载体PCR和测序结果与预期结果一致,经包装产生的病毒滴度为1.58×1011U/ml。NF-κB-p65-sh RNA腺病毒感染食蟹猴子宫内膜细胞后,实验组子宫内膜细胞凋亡蛋白的表达水平明显高于阴性对照组(P<0.05);实验组凋亡细胞较对照组明显增多,进入细胞分裂期细胞数明显少于对照组(P<0.05)。结论携带NF-κB基因的腺病毒体外感染食蟹猴子宫内膜细胞后可加速子宫内膜细胞的凋亡,并抑制子宫内膜细胞的增殖,为后期该基因在食蟹猴子宫内膜异位症模型上的研究提供理论基础。Objective To assess the effect of a high specific adenovirus vector-mediated sh RNA targeting nuclear factor-κB（NF-κB） on cell proliferation of the endometrium of Macaca fascicularis. Methods The adenoviral vector NF-κB-p65-sh RNA and the empty vector were separately trasnfected in cultured endometrial cells of Macaca fascicularis. The changes in the expression of the target gene protein and apoptotic proteins, cell proliferation, and cell cycle distribution were observed after the transfection. Results Compared with the control cells, infection of the endometrial cells with the NF- κB- p65- sh RNA adenovirus significantly increased the expression levels of apoptotic proteins, promoted apoptosis of the endometrial cells,and reduced the cells in division stage. Conclusions NF- κB- p65 sh RNA adenovirus can effectively promote apoptosis of endometrial cells and inhibit the proliferation of endometrial cells of Macaca fascicularis.

关 键 词：子宫内膜异位症 RNA干扰 NF-κB基因 腺病毒 食蟹猴 

分 类 号：R711.71[医药卫生—妇产科学]