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(Adh S100A9)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(vimentin,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(P<0.05)和Vim升高(P<0.01);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(P<0.01),并且入核增多(P<0.01),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(epithelial-mesenchymal transition,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。
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 signaling pathway, respectively. The results showed that S100A9 over-expression promoted the prolifera- tion and migration of Hela cells, and induced a morphology transition from a round cobblestone-like phenotype to a slender and fusiform appearance, also led to reduction of E-cadherin (P〈0.05) and increase of Vimentin (P〈0.01). Moreover, the levels of β-catenin, c-myc, Snail and Twist in S 100A9 over-expressed Hela cells were also increased. The results demonstrated that S 100A9 could promote the proliferation, migration and epithelial-mesenchymal tran-sition (EMT) of Hela cells and also activated the WntJl3-catenin signaling pathway. The mechanisms that S 100A9 increased the cell proliferation and migration of Hela cell line might be associated with its promoting effect on EMT and the Wnt/β-catenin signaling pathway.
出处
《中国细胞生物学学报》
CAS
CSCD
2015年第12期1632-1638,共7页
Chinese Journal of Cell Biology
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
S 100A9
cervical carcinoma
cell proliferation
cell migration
epithelial-mesenchymal transi-tion
Wnt/β-catenin signaling pathway