S100A6促宫颈癌细胞增殖、迁移及其可能机制研究

发布时间：2018-02-28 07:41

  本文关键词： 宫颈癌 SA 上皮间质转化 PIK-Akt信号通路　出处：《中国生物工程杂志》2017年02期 　论文类型：期刊论文

【摘要】：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。
[Abstract]:Objective: to investigate the effect of S100A6 on the proliferation and migration of human cervical cancer cell line HeLa and SiHa and its mechanism. Methods: the basic expression of S100A6 mRNA in cervical cancer cell line HeLa La SiHa and CaSki was detected by quantitative polymerase chain reactionqPCRs. Then the recombinant adenovirus AdS100A6 and AdsiS100A6 were used to test the proliferation of HeLa and SiHa cells by Western blot. The ability of cell migration was detected by scratch healing test and the levels of E-cadherin N-cadherin N-cadherin N-cadherin and p-Akt protein were detected by blot and E-cadherin E-cadherin N-cadherin N-cadherin N-cadherin and p-Akt protein levels in epithelium epithelial-mesenchymal transition. Semi-quantitative reverse transcription and polymerase chain reactionation polymerase chain reaction (RT-PCR) was used to detect the expression of the downstream target gene Snail-Twist in the PI3K-Akt signaling pathway. Results: compared with the control group, the HeLa cells in the AdS100A6 group were significantly higher in ODS-492) value and scratch healing rate than those in the control group. With the decrease of E-cadherin and the increase of N-cadherin, the number of SiHa cells in AdsiS100A6 group decreased significantly at 5 days and the rate of scratch healing at 3 days, and the E-cadherin increased and N-cadherin decreased, at the same time, the level of p-Akt protein increased after S100A6 was up-regulated in HeLa cells. The downstream target gene Snail and Twist expression were also up-regulated. Conclusion: S100A6 can enhance the proliferation and migration of cervical cancer cells, and its mechanism may be related to the activation of EMT and PI3K-Akt signaling pathway.
【作者单位】： 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;
【基金】：重庆市研究生科研创新资助项目(CYS15133)
【分类号】：R737.33

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