下调p21活化蛋白激酶2表达对人乳腺癌细胞增殖和凋亡的影响  

Effects of p21-activated protein kinase 2 down-regulation on proliferation and apoptosis of human breast cancer cells

作　　者：李翔[1] 张晓艳[1] 赵继敏[1] 刘康栋[1] 赵明耀[1] 董子明[1] 

机构地区：[1]郑州大学基础医学院病理生理学教研室,河南郑州450001

出　　处：《中国病理生理杂志》2014年第6期975-981,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31301144);国家教育部博士学科点专项科研基金(No.20134101120011);河南省教育厅科学技术研究重点项目(No.13A310564;No.14A310010)

摘　　要：目的:应用RNA干扰技术下调人乳腺癌MCF-7细胞中p21活化蛋白激酶2(PAK2)的表达,观察其对MCF-7细胞增殖和凋亡的影响。方法:构建针对PAK2基因的短发卡RNA(shRNA)慢病毒表达载体并包装病毒颗粒,感染MCF-7细胞,筛选获得稳定细胞株。Western blotting检测细胞中PAK2蛋白表达水平的改变;CellTiter96 AQueous法和软琼脂集落形成实验检测MCF-7细胞体外增殖能力的改变;流式细胞术检测十字孢碱诱导的MCF-7细胞凋亡的变化。结果:MCF-7细胞的PAK2基因被沉默后,PAK2蛋白表达量明显下降(P<0.01),与阴性对照组相比,sh-PAK2组细胞生长明显减缓(P<0.01),克隆形成的数目和大小明显下降(P<0.01),十字孢碱诱导的细胞凋亡率显著增加(P<0.01)。结论:通过RNA干扰技术下调PAK2的表达,可抑制MCF-7细胞的生长和增殖,并增强其对化疗药物诱导的细胞凋亡的敏感性。PAK2可能是潜在的乳腺癌治疗的新靶点。AIM：To study the effect of p21-activated protein kinase 2 （PAK2） knockdown by RNA interfer-ence on the proliferation and apoptosis of human breast cancer cells .METHODS：The short hairpin RNA （ shRNA） targeting PAK2 gene was designed and used for packing lentivirus in 293T cells.Human breast cancer MCF-7 cells were infected by the virus particles and PAK2 knockdown stable cell line was established by puromycin selection .The knockdown efficiency was assessed by Western blotting .The proliferation ability of MCF-7 cells was evaluated by CellTiter 96 AQueous and anchorage-independent growth assays .The cell apoptosis induced by staurosporine was detected by flow cytometry . RESULTS：The protein level of PAK2 was significantly suppressed after silencing of PAK2 gene in MCF-7 cells （ P〈0.01）.Furthermore, knockdown of PAK2 caused remarkable inhibition of the cell proliferation and colony formation （P〈0.01）.Staurosporine induced more apoptosis in the PAK2 knockdown cells compared with the control cells （P〈0.01）. CONCLUSION：Knockdown of PAK2 inhibits the proliferation of MCF-7 cells and increases the sensitivity of chemothera-peutic druginduced cell apoptosis , suggesting that PAK2 might be a new therapeutic target in breast cancer treatment .

关 键 词：p21活化蛋白激酶2 乳腺肿瘤 RNA干扰 细胞增殖 细胞凋亡 

分 类 号：R737.9[医药卫生—肿瘤]