蒸馏水促进人晶状体上皮细胞(SRA01/04)凋亡并抑制其增殖、迁移、侵袭及转分化能力研究

Effects of distilled water on apoptosis,proliferation,migration,invasion,and transdifferentiation of human lens epithelial cells SRA01/04

目的探究蒸馏水(DW)对人晶状体上皮细胞SRA01/04凋亡、增殖、迁移、侵袭及转分化的影响。方法将人晶状体上皮细胞系SRA01/04细胞随机分为5组:对照组(不作处理)、BBS组(BBS处理3 min)、DW 30 s组(DW处理30 s)、DW 1 min组(DW处理1 min)、DW 2 min组(DW处理2 min)、DW 3 min组(DW处理3 min)。除对照组外,其余组加入2 mL细胞培养液终止作用,继续培养24 h。流式细胞仪检测各组细胞凋亡情况;5-乙炔基-2’-脱氧尿苷和CCK-8实验检测各组细胞增殖情况;Western blot检测各组凋亡相关蛋白Bax、Bcl-2、Caspase-9及转分化相关蛋白α-平滑肌肌动蛋白(α-SMA)、N-钙黏蛋白(N-cadherin)、纤连蛋白(Fibronectin)、波形蛋白(Vimentin)的表达;细胞划痕实验检测各组细胞迁移情况;Transwell细胞迁移实验检测各组细胞迁移及侵袭情况。使用单因素方差分析(ANOVA)进行多组间的比较,组间两两比较采用Tukey检验。结果随着DW作用时间的逐渐增加,SRA01/04细胞凋亡率逐渐增加;SRA01/04细胞的增殖、迁移及侵袭能力受到明显抑制;SRA01/04细胞促凋亡相关蛋白Bax、Caspase-9表达逐渐增加,抗凋亡蛋白Bcl-2表达逐渐降低;转分化相关蛋白α-SMA、N-cadherin、Fibronectin及Vimentin表达逐渐降低,SRA01/04细胞EMT作用受到抑制。结论DW可以促进SRA01/04细胞凋亡,抑制SRA01/04细胞增殖、迁移、侵袭及转分化能力。

Objective To investigate the effects of distilled water(DW)on apoptosis,proliferation,migration,invasion and transdifferentiation of human lens epithelial cells SRA01/04.Methods The cultured SRA01/04 cells were divided into 5 groups:control group(no treatment),BBS group(BBS treatment for 3 min),30 s DW group(DW treatment for 30 s),1 min DW group(DW treatment for 1 min),2 min DW group(DW treatment for 2 min),and 3 min DW group(DW treatment for 3 min).Except for the control group,the other groups were added with 2 mL of cell culture medium and continued to culture for 24 hours.The cell apoptosis was detected by flow cytometry.The cell proliferation was detected by 5-acetylidene-2’-deoxyuridine and CCK-8.The expression levels of apoptosis-related proteins such as B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(BAX)and Caspase-9,and transdifferentiation-related proteins such asα-smooth muscle actin(α-SMA),N-cadherin,Fibronectin and Vimentin were measured by Western blot.Cell migration was detected by cell scratch assay.Cell migration and invasion were detected by Transwell assay.One-way analysis of variance was used for multi-group comparison,and the Tukey test was used for comparison between groups.Results With the increase in DW treatment time,the cell apoptosis increased;the cell proliferation,migration and invasion were significantly inhibited;the expression levels of pro-apoptosis-related proteins Bax and Caspase-9 increased,while the expression level of anti-apoptosis-related protein Bcl-2 decreased;the expression levels of transdifferentiation-related proteinsα-SMA,N-cadherin,Fibronectin and Vimentin decreased,and the epithelial-mesenchymal transition of SRA01/04 cells was inhibited.Conclusion DW can promote apoptosis of SRA01/04 cells and inhibit proliferation,migration,invasion and transdifferentiation of SRA01/04 cells.