S100A6促宫颈癌细胞增殖、迁移及其可能机制研究

Effects of S100A6 on Proliferation and Migration of Human Cervical Cancer Cells and Its Mechanism

目的:探讨S100A6对人宫颈癌细胞系HeLa和SiHa增殖、迁移的影响及其机制。方法:首先采用定量聚合酶链反应(quantitative polymerase chain reaction,qPCR)检测宫颈癌细胞HeLa、SiHa和CaSki中S100A6 mRNA的基础表达,再分别采用重组腺病毒AdS100A6和AdsiS100A6干预HeLa和SiHa细胞,Western blot验证腺病毒感染是否成功;MTT法检测细胞增殖能力,划痕愈合试验检测细胞迁移能力,Western blot检测上皮间质转化(epithelial-mesenchymal transition,EMT)指标E钙粘蛋白(E-cadherin,E-cad)、N钙粘蛋白(N-cadherin,N-cad)及p-Akt的蛋白水平,半定量反转录聚合酶链反应(reverse transcription and polymerase chain reaction,RT-PCR)检测PI3K-Akt信号通路下游靶基因Snail、Twist的表达。结果:与对照组相比,AdS100A6组的HeLa细胞3天时的OD_(492)值和划痕愈合率均明显升高,并伴随E-cadherin降低和N-cadherin升高;而AdsiS100A6组的SiHa细胞5天时的OD_(492)值和3天时的划痕愈合率明显降低,并伴随E-cadherin升高和N-cadherin降低;同时,在HeLa细胞中上调S100A6后p-Akt蛋白水平增加,该通路的下游靶基因Snail和Twist表达也明显上调。结论:S100A6可以增强宫颈癌细胞的增殖和迁移能力,其机制可能涉及EMT和PI3K-Akt信号通路的激活。

Objective： To investigate the effects of S100A6 on cell proliferation and migration of cervical cancer cells and explore its mechanism. Methods： Quantitative polymerase chain reaction（ qPCR） was used to detect the basic expression of S100A6 in HeLa,SiHa and CaSki cells; HeLa and SiHa cells were treated with recombinant adenoviruses AdS100A6 and AdsiS100A6 to upregulate and downregulate S100A6 expression respectively,Western blot was used to verify the effects of virus infection. MTT method was used to detect cell proliferation ability; wound healing assay was used to detect cell migration ability; Western blot was used to detect the level of E-cadherin,N-cadherin and p-Akt; reverse transcription and polymerase chain reaction（ RTPCR） was used to detect the level of Snail and Twist,the downstream target genes of PI3K-Akt pathway.Results： In AdS100A6 group（ HeLa cells）,the OD492 value and wound healing rate at the 3rd day were increased,N-cadherin was increased,but E-cadherin was decreased; In AdsiS100A6 group（SiHa cells）,the OD492 value at the 5th day and wound healing rate at the 3rd day were decreased,N-cadherin was decreased,but E-cadherin was increased; At the same time,in AdS100A6 group,p-Akt was increased,Snail and Twist were also increased obviously. Conclusion： S100A6 promotes cervical cancer cells proliferation and migration and the effect may be mediated by inducing epithelial-mesenchymal transition and activating PI3K / Akt signaling pathway.