ANO1稳定高表达对Hep-2细胞株生物学特性的影响  

Effects of stable ANO1 overexpression on biological behaviors of human laryngeal squamous cell carcinoma Hep-2 cells in vitro

作　　者：李雅冬[1] 张劲松[1] 杨凯[1] 张福军[1] 陈睿[1] 陈丹[1] 

机构地区：[1]重庆医科大学附属第一医院颌面外科,重庆400016

出　　处：《南方医科大学学报》2014年第2期251-255,共5页Journal of Southern Medical University

基　　金：重庆市自然科学基金(cstc2012jjA10039);重庆市卫生局医学科研计划项目(2011-2-013);重庆市教委科学技术研究项目(KJ130318)

摘　　要：目的检测ANO1高表达对Hep-2细胞株生物学特性的影响。方法利用ANO1稳定高表达的Hep-2细胞株先后进行流式细胞仪,软琼脂细胞生长实验,体外划痕愈合实验,SiRNA实验,SiRNA沉默ANO1的体外划痕愈合实验,DIDS阻断氯离子通道实验,以明确ANO1高表达对Hep-2细胞株生长、迁移及侵袭的影响。结果流式细胞仪检测细胞周期,结果显示实验组和空白组的G0/G1期比率差异无显著性(P>0.05);软琼脂细胞生长实验结果显示实验组和对照组差异无显著性(P>0.05);体外划痕愈合实验结果显示两者差异有显著性(P<0.05),表明ANO1高表达加快了Hep-2细胞的迁移速度;SiRNA沉默ANO1的体外划痕愈合实验结果显示两者差异有显著性(P<0.05),表明沉默ANO1可减缓Hep-2细胞的迁移速度。DIDS阻断氯离子通道实验结果显示两者差异有显著性(P<0.05),表明DIDS能有效的阻断ANO1的氯离子通道活性,并且减缓Hep-2细胞的迁移速度,再一次证明ANO1参与了Hep-2细胞的迁移活动。结论 ANO1高表达并未改变癌细胞的增殖速度,但却大大增加了头颈鳞癌细胞的移动能力,有望成为治疗头颈鳞癌的新靶点。Objective To detect the effects of ANO1 overexpression on the biological behaviors of human laryngeal squamous cell carcinoma Hep-2 cells. Methods A Hep-2 cell line stably overexpressing ANO1 were examined with flow cytometry, soft agar assay, wound healing assay, siRNA experiments, and chloride channel block with DIDS to observe the effect of ANO1 overexpression on the growth, migration and invasion of the cells. Results Flow cytometry revealed a comparable cell percentage in G0/G1 phase between ANO1-overexpressing cells and the control cells （P＆gt;0.05）. The two cells showed no significant difference in soft agar assay （P＆gt;0.05）, but in wound healing experiments, ANO1-overexpressing cells showed significantly accelerated migration （P＆lt;0.05）, whereas siRNA-mediated silencing of ANO1 significantly inhibited the cell migration （P＆lt;0.05）. Treatment with DIDS resulted in an effective block of the ANO1 chloride channel activity and obviously decreased the migration speed of Hep-2 cells. Conclusion ANO1 overexpression does not significantly affect the proliferation of cancer cells, but can enhance the migration ability of head and neck squamous cell carcinoma, suggesting the value of ANO1 as a new gene therapy target for head and neck squamous cell carcinoma.

关 键 词：头颈鳞癌 ANO1 高表达 氯离子通道 生物学特性 HEP-2细胞 ANO1 

分 类 号：R739.91[医药卫生—肿瘤]