替米沙坦通过Wnt/β-catenin信号通路影响非小细胞肺癌A549细胞增殖、迁移和凋亡的研究

Telmisartan affects proliferation,migration and apoptosis of non-small cell lung cancer cell A549 through the Wnt/β-catenin signaling pathway

目的研究替米沙坦对非小细胞肺癌A549细胞增殖、迁移和凋亡的影响,并探讨其机制。方法体外培养非小细胞肺癌细胞株A549,采用CCK-8法检测不同浓度替米沙坦对A549细胞增殖活性的影响,以克隆形成实验检测不同浓度替米沙坦处理下A549的存活分数,以划痕实验检测不同浓度替米沙坦对A549细胞迁移能力的影响,Hoechst染色检测不同浓度替米沙坦对A549凋亡的影响,用Western blotting检测β-肌动蛋白(β-actin)、增殖细胞核抗原(PCNA)、Bax、Bcl-2、Wnt-3a、β-连环蛋白(β-catenin)、丝氨酸蛋白激酶3β(p-GSK-3β)、糖原合成酶激酶-3β(GSK-3β)、c-myc等蛋白的表达。结果不同浓度替米沙坦处理抑制了A549细胞增殖活性、克隆形成率和迁移,减少PCNA表达,并且存在浓度依赖性。替米沙坦处理促进了A549细胞的凋亡,显著增加促凋亡蛋白Bax和减少抗凋亡蛋白Bcl-2的表达。替米沙坦处理后A549细胞的Wnt-3a、β-catenin、c-myc、p-GSK-3β的表达减少,GSK-3β的表达增加。结论替米沙坦可能在非小细胞肺癌A549细胞的增殖、迁移和凋亡中起到作用,抑制Wnt/β-catenin信号通路可能是其中的一个机制。

Objective To investigate the effects of telmisartan on the proliferation,migration and apoptosis of non-small cell lung cancer A549 and the mechanism of regulating Wnt signaling pathway.Methods Non-small cell lung cancer cell line A549 was cultured in vitro.Cell counting kit-8(CCK-8)assay was used to detect the effect of telmisartan at different concentrations on the proliferative activity of A549 cells.The survival fraction of A549 treated with different concentrations of telmisartan was determined by colony-formation assay.The effect of telmisartan at different concentrations on the migration ability of A549 cells was examined in the wounding healing assay.Hoechst staining was used to detect the effects of telmisartan at different concentrations on the apoptosis of A549.Western bloting was used to detect the expressions ofβ-actin,proliferating cell nuclear antigen(PCNA),Bax,Bcl-2,Wnt-3 a,Beta-catenin(β-catenin),serine protein kinase 3β(p-GSK-3β),glycogen synthase kinase-3β(GSK-3β)and c-myc.Results Different concentrations of telmisartan treatment inhibited the proliferation activity,colony-formation rate and migration of A549 cells,and reduced the expression of PCNA in a concentration-dependent manner.Telmisartan treatment promoted the apoptosis of A549 cells,significantly increased the expression of pro-apoptotic protein Bax and decreased the expression of antiapoptotic protein Bcl-2.The expression levels of Wnt-3 a,β-catenin,p-GSK-3β,and c-myc in A549 cells increased after treatment with telmisartan,while the expression levels of GSK-3βdecreased.Conclusion Telmisartan may play a role in the proliferation,migration and apoptosis of non-small cell lung cancer A549 cells,and inhibiting the Wnt/β-catenin signaling pathway may be one of the mechanisms.