YAP干扰腺病毒载体对宫颈癌增殖、迁移及侵袭的影响  

Effect of adenovirus-mediated silencing of YAP on the proliferation, migration,and invasion of cervical carcinoma

作　　者：王红[1] 王恳[1] 焦庆昉[1] 杨万全[1] 张镇[1] 李婕婷[1] 吴扶生 王瑾 WANG Hong;WANG Ken;JIAO Qingfang;YANG Wanquan;ZHANG Zhen;LI Jieting;WU Fusheng;WANG Jin(Department of Oncology,Sichuan Hospital of Integrated Traditional and Western Medicine,Chengdu,Sichuan 610041,China;Department of Hematology,The 989th Hospital of the Joint Logistic Support Force of the People's Liberation Army,Luoyang,Henan 471003,China)

机构地区：[1]四川省中西医结合医院肿瘤科,四川成都610041 [2]中国人民解放军联勤保障部队第九八九医院血液科,河南洛阳471003

出　　处：《安徽医药》2021年第6期1136-1139,I0004,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探索YAP蛋白对宫颈癌细胞的恶性增殖、迁移及侵袭性的作用。方法分别以腺病毒靶向YAP干扰载体(pAd-si-YAP)、重组空载体腺病毒(pAd-mock)或磷酸缓冲盐溶液(PBS)感染CaSki细胞。采用定量聚合酶链反应(qPCR)、蛋白质印迹法(Western blotting)、四甲基偶氮唑盐微量酶反应比色法(MTT法)、流式细胞法、划痕试验以及细胞侵袭试验分别分析感染后CaSki细胞的YAP mRNA表达、YAP蛋白表达、细胞增殖活力、细胞周期及凋亡、细胞迁移能力以及细胞侵袭能力。结果qPCR结果显示Ad-si-YAP感染CaSki细胞的YAP mRNA表达明显低于pAd-mock感染的CaSki细胞及PBS处理的CaSki细胞[(22.01±0.24)比(80.12±0.31)、(84.18±0.22),P<0.05],表明Ad-si-YAP可以抑制YAP基因转录。蛋白质印迹法结果显示Ad-si-YAP感染CaSki细胞的YAP蛋白表达低于pAd-mock感染的CaSki细胞及PBS处理的CaSki细胞[(0.6±0.018)比(1.5±0.031)、(1.8±0.27),P<0.05]。MTT结果显示24 h后pAd-mock感染的CaSki细胞及PBS处理的CaSki细胞增殖明显快于Ad-siYAP感染CaSki细胞(P<0.05)。流式细胞结果显示通过沉默YAP后,CaSki细胞周期停滞于G0/G1期明显增加(P<0.05),同时CaSki细胞增多(P<0.01)。划痕试验发现沉默YAP后CaSki细胞的迁移能力下降[(40.01±0.16)比(81.02±0.22)、(86.04±0.31),P<0.05],说明YAP可以促进CaSki细胞迁移。细胞侵袭试验发现沉默YAP后CaSki细胞的侵袭能力下降[(120±4)比(640±3)、(680±4),P<0.05]。结论YAP蛋白的表达可导致宫颈癌细胞的恶性增殖、迁移及侵袭,是应用于宫颈癌综合治疗的一个潜在靶点。Objective To explore the effect of adenovirus-mediated YAP-silencing system(pAd-si-YAP)on the malignant proliferation,migration,and invasion of cervical carcinoma.Methods pAd-si-YAP,mock-recombinant adenovirus(pAd-mock),and phosphate buffer saline(PBS)were transfected into CaSki cell lines.The YAP mRNA/protein expression level,the cell proliferative potential,the cell cycle and apoptosis,and the migration/invasive potential of the transfected CaSki cell lines were determined by using quantitative polymerase chain reaction(qPCR),Western blotting,MTT assay,flow cytometry,wound healing assay,and Transwell assay,respectively.Results In this experiment,qPCR indicated that CaSki cells transfected with Ad-si-YAP showed a significantly lower YAP mRNA expression level compared to those transfected with pAd-mock or treated with PBS[(22.01±0.24)vs.(80.12±0.31),(84.18±0.22),P<0.05].Western blotting indicated that CaSki cells transfected with Ad-si-YAP showed a significantly lower YAP protein expression level compared to those transfected with pAd-mock or treated with PBS[(0.6±0.018)vs.(1.5±0.031),(1.8±0.27),P<0.05].MTT assay indicated that CaSki cells transfected with p Ad-mock or treated with PBS proliferated more rapidly compared to those transfected with Adsi-YAP(P<0.05).Flow cytometry indicated that Yap silencing caused more CaSki cells to arrest at the G0/G1 phase(P<0.05)and resulted in a higher cell count(P<0.01).Wound healing assay revealed a decrease in the migration potential of the CaSki cells after YAP silencing[(40.01±0.16)vs.(81.02±0.22),(86.04±0.31),P<0.05].Transwell assay revealed a decrease in the invasive potential of the CaSki cells after YAP silencing[(120±4)vs.(640±3),(680±4),P<0.05].Conclusions The expression of YAP protein can induce malignant proliferation,migration,and invasion of cervical carcinoma cells,and it is a potential target for the comprehensive treatment of cervical carcinoma.

关 键 词：子宫肿瘤 YAP Hippo信号通路 CASKI细胞 恶性增殖 迁移 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]