膜蛋白AN01过表达对人喉鳞状细胞癌Hep-2细胞株生物学特性的影响

目的 研究膜蛋白ANO1过表达对人喉鳞状细胞癌Hep-2细胞增殖、剥脱、伸展和迁移的影响。方法 以ANO1稳定过表达的Hep-2细胞株作为实验组,空白质粒转染的Hep-2细胞作为对照组,采用MTT法检测细胞增殖活性,细胞剥脱实验检测细胞剥脱能力,细胞伸展实验检测细胞伸展能力,Boyden小室侵袭实验、体外划痕愈合实验和尼氟灭酸阻断氯离子通道实验检测细胞迁移能力。结果 MTT法检测结果显示,实验组与对照组的光密度值差异无统计学意义(P=0.62)。细胞剥脱实验和细胞伸展实验结果显示,实验组的细胞剥脱百分比(P＜0.0001)和伸展百分比(P＜0.0001)明显大于对照组。Boyden小室侵袭实验结果显示,实验组的穿膜细胞百分比明显大于对照组(P＜0.0001);体外划痕愈合实验结果显示,实验组的划痕面积百分比明显小于对照组(P＜0.0001);尼氟灭酸阻断氯离子通道实验结果显示,实验组的划痕面积百分比明显大于对照组(P＜0.0001)。结论 ANO1过表达并未加快癌细胞的增殖速度,但却大大增加了头颈鳞癌细胞的移动、伸展和剥脱能力。

Effects of Membrane Protein ANO1 Stable Overexpression on Laryngocarcinoma Hep-2 Cells

Objective To explore the effects of ANO1 overexpression on the proliferation,detachment,spreading,and migration of laryngocarcinoma Hep-2 cell line.Methods ANO1-overexpressing Hep-2 cell line was selected as the assay group,and Hep-2 cell line with empty plasmid was selected as the control group.MTT assay was used to detect the proliferation abilities of Hep-2 cells in both two groups.Cell detachment assay and spreading assay were used to detect the detachment and spreading abilities of Hep-2 cells.Boyden chamber invasion assay,wound healing assay in vitro,and niflumic acid block chloride channel were used to detect the migration abilities of Hep-2 cells.All data were analyzed by SPSS 10.0 software package.Results Cell proliferation assay by MTT showed that,compared with the control group,the optical density value of assay group was not significantly different(P=0.62).The results of cell detachment assay and cell spreading assay showed the cell detachment rates and cell spreading rates in assay group were significantly higher than those in control group(P＜0.0001).The results of Boyden chamber invasion assay showed the percentages of cells migrating through the membrane in assay group were significantly higher than those in control group(P＜0.0001).The results of in vitro wound healing experiments showed the wound area rate in assay group was significantly lower than that in control group(P＜0.0001).The results of niflumic acid blocking chloride channel experiments showed the wound area rates in assay group were significantly higher than those in control group(P＜0.0001).Conclusion ANO1 overexpression does not remarkably alter the proliferation rate of cancer cells,but increases the migration,spreading,and detachment capacities of head and neck squamous cell carcinoma.