过表达同源盒A5对人甲状腺癌细胞增殖的影响及作用机制
Effect of overexpression of HOXA5 on proliferation of human thyroid cancer cells and its mechanism
田荣英 1王缚鲲 1安黎云 1李芳 1张会峰 1郑运周 1冉向阳1
作者信息
- 1. 中国人民解放军联勤保障部队第980医院检验科,石家庄 050081
- 折叠
摘要
目的:探讨同源盒A5(HOXA5)对人甲状腺癌 TPC1 细胞生物学功能的影响,并探讨其可能的作用机制.方法:采用 RT-qPCR和免疫印迹分析人甲状腺癌 TPC1 细胞中的HOXA5 mRNA和蛋白表达水平.采用慢病毒转染技术将阴性对照质粒(OE-NC)和HOXA5过表达质粒(OE-HOXA5)转染至TPC1细胞,并验证转染效率;CCK-8法、克隆形成实验和流式细胞术分别检测细胞增殖、克隆形成及细胞周期;免疫印迹检测增殖相关蛋白及AKT通路相关蛋白的表达水平.结果:与甲状腺滤泡上皮细胞相比,甲状腺癌TPC1细胞中的HOXA5 mRNA和蛋白表达水平明显降低.上调HOXA5表达后,TPC1细胞的增殖活性和克隆形成能力明显受到抑制,细胞在G0/G1期的细胞百分比显著增加,但在S期的细胞百分比下降;且增殖相关蛋白Ki67、c-Myc及AKT信号通路关键蛋白p-GSK3β、p-AKT表达水平明显下调.结论:HOXA5在甲状腺癌TPC1细胞中表达下调,且过表达HOXA5基因可抑制甲状腺癌细胞增殖和克隆形成能力,其机制可能涉及AKT信号通路的激活.
Abstract
Objective:To investigate the effect of homeobox A5(HOXA5)on the biological function of human thyroid cancer TPC1 cells and its possible mechanism.Methods:The expression of HOXA5 mRNA and protein in TPC1 cells was analyzed by RT-qPCR and Western blotting.Negative control plasmid(OE-NC)and HOXA5 overexpression plasmid(OE-HOXA5)were transfected into TPC1 cells using lentiviral transduction and the transfection efficiency was verified.Cell proliferation,colony forming ability and cell cycle were detected by CCK-8,colony formation assay and flow cytometry,respectively.The expression of proliferation related proteins and AKT pathway related proteins were detected by Western blotting.Results:The expression levels of HOXA5 mRNA and protein in thyroid cancer cells were significantly lower than that in thyroid follicular epithelial cells.After up-regulation of HOXA5 expression,the proliferation and clonogenesis of TPC1 cells were significantly inhibited,and the percentage of cells in G0/G1 phase was significantly increased,but the percentage of cells in S phase was decreased.The expression levels of proliferation-related proteins Ki67,c-Myc,as well as key proteins in the AKT signaling pathway,p-GSK3β,and p-AKT were significantly down-regulated.Conclusion:HOXA5 is down-regulated in thyroid cancer,and overexpression of HOXA5 gene can inhibit the proliferation and clonogenesis of thyroid cancer cells,and its mechanism may involve the activation of AKT signaling pathway.
关键词
同源盒A5/甲状腺癌/细胞增殖/细胞周期/AKT信号通路Key words
HOXA5/thyroid carcinoma/cell proliferation/cell cycle/AKT signaling pathway引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据