S100A9对宫颈癌Hela细胞的增殖和迁移的影响及其机制探讨.pdfVIP

中国细胞生物学学报 Chinese Journal of Cell Biology 2015, 37(12): DOI: 10.11844/cjcb.2015.12.0246 收稿日期: 2015-07-26 接受日期: 2015-10-26 *通讯作者。Tel: E-mail: zhoulan0111@ Received: July 26, 2015 Accepted: October 26, 2015 *Corresponding author. Tel: +86 E-mail: zhoulan0111@ S100A9对宫颈癌Hela细胞的增殖和 迁移的影响及其机制探讨 李雪茹 孙 晖 查 何 段 亮 李 欢 袁世梅 李爱芳 谷 月 周 兰* (重庆医科大学检验医学院, 临床检验诊断学教育部重点实验室, 重庆 400016) 摘要 为初步探讨S100A9在宫颈癌中的生物学作用, 利用携带有人S100A9基因的腺病毒 (AdhS100A9)感染于低表达S100A9的宫颈癌Hela细胞, 通过MTT法检测细胞增殖能力, 划痕愈合试 验和Transwell试验检测细胞迁移能力, 通过倒置显微镜观察细胞形态变化, 采用RT-PCR和蛋白质 印迹法检测上皮细胞标志蛋白E-钙黏着蛋白(E-cadherin, E-cad)、间质细胞标志蛋白波形蛋白(vi- mentin, Vim)及Wnt/β-联蛋白(β-catenin, β-cat)信号通路相关分子的表达。结果显示, 与对照组相比, 过表达S100A9的Hela细胞增殖活性增强、迁移率增高, 细胞形态由“铺路石”样向“梭形”转变, 排 列紊乱, 伴随E-cad降低(P0.05)和Vim升高(P0.01); 同时, 过表达S100A9的Hela细胞中Wnt/β-cat 信号通路的关键分子β-联蛋白水平增加(P0.01), 并且入核增多(P0.01), 该通路的下游靶基因c- myc、Snail及Twist表达也明显上调。该研究结果提示, S100A9促进宫颈癌Hela细胞的增殖、迁移和 上皮间质转化(EMT), 激活Wnt/β-cat信号通路; S100A9促进增殖和迁移作用的机制可能与其促进 EMT和激活Wnt/β-cat信号通路相关。 关键词 S100A9; 宫颈癌; 增殖; 迁移; 上皮间质转化; Wnt/β-联蛋白信号通路 The Effects of S100A9 on Proliferation and Migration of Human Squamous Cervical Carcinoma Hela Cell Line and Its Mechanisms Li Xueru, Sun Hui, Zha He, Duan Liang, Li Huan, Yuan Shimei, Li Aifang, Gu Yue, Zhou Lan* (Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China) Abstract To investigate the role of S100A9 in human squamous cervical carcinoma, the cervical carci- noma cells (Hela cell line) were infected with recombinant adenovirus expressing S100A9. Then, MTT was used to detect cell proliferation; Wound-healing test and Transwell assay were used to determine cell migration; Inverted microscope was used to observe the morphologic changes; Reverse transcription-PCR (RT-PCR) and Western blot were utilized to detect the mRNA or protein levels of E-Cadherin, Vimentin and the relevant molecules of Wnt/ β-catenin signalin