麦冬皂苷B抑制人食管鳞状细胞癌细胞系EC9706增殖和迁移

Ophiopogonin B inhibits proliferation and migration of human esophageal squamous cell carcinoma cell line EC9706

目的探讨麦冬皂苷B(OP-B)对食管鳞状细胞癌细胞系(EC9706)增殖、迁移和凋亡的影响,分析其机制是否与调控唐氏综合征细胞黏附分子反义1(DSCAM-AS1)和miR-199a-3p表达有关。方法将EC9706细胞分为对照组、低、中、高OP-B组、si-NC组、si-DSCAM-AS1组、(pcDNA-DSCAM-AS1+OP-B)组。集落形成实验和CCK-8法检测EC9706增殖。流式细胞测量术、划痕愈合实验分析EC9706细胞凋亡率和迁移能力。RT-qPCR检测DSCAM-AS1和miR-199a-3p表达量。双荧光素酶报告基因实验确定DSCAM-AS1和miR-199a-3p靶向关系。结果OP-B处理将降低EC9706活力、集落形成数、迁移距离和DSCAM-AS1表达量(P<0.05),增加细胞凋亡率和miR-199a-3p表达量(P<0.05)。DSCAM-AS1与miR-199a-3p存在直接相互作用。干扰DSCAM-AS1表达后,EC9706活力、克隆形成数、迁移距离显著降低(P<0.05),细胞凋亡率显著增加(P<0.05)。过表达DSCAM-AS1可逆转OP-B直接处理对EC9706细胞增殖、凋亡和迁移的影响(P<0.05)。结论OP-B可抑制食管鳞状细胞癌细胞增殖和迁移,诱导细胞凋亡,其机制可能是通过下调DSCAM-AS1/miR-199a-3p途径实现的。

Objective To explore the effect of ophiopogonin B(OP-B)on the biological behavior of esophageal squamous cell carcinoma cell line(EC9706)and its mechanism related to the regulation of Down syndrome cell adhesion molecule antisense 1(DSCAM-AS1)and miR-199a-3p expression.Methods EC9706 cells were divided into control group,experiment groups with low,medium and high dosage of OP-B,si-NC group,si-DSCAM-AS1 group and(pcDNA-DSCAM-AS1+OP-B)group.Colony formation and CCK-8 method were used to detect EC9706 proliferation.Flow cytometry and scratch healing test were used to analyze the apoptosis rate and migration of EC9706 cells.RT-qPCR was performed to detect the expression of DSCAM-AS1 and miR-199a-3p.The targeting relationship between DSCAM-AS1 and miR-199a-3p was verified using dual luciferase reporter gene experiment.Results OP-B reduced EC9706 viability,colony formation counting,migration distance and DSCAM-AS1 expression(P<0.05),increased cell apoptosis rate and miR-199a-3p expression(P<0.05).DSCAM-AS1 interacted directly with miR-199a-3p.After interference to DSCAM-AS1 expression,the cell viability,colony formation and migration distance of EC9706 cells were significantly reduced(P<0.05),while apoptosis rates were significantly increased(P<0.05).Over-expressionof DSCAM-AS1 antagonized the effects of OP-B on the proliferation,apoptosis and migration of EC9706 cells(P<0.05).Conclusions OP-B inhibits proliferation and migration of esophageal squamous cell carcinoma cells and induces cell apoptosis.The mechanism is potentially explained by down-regulating DSCAM-AS1/miR-199a-3p pathway.