miR-454-3p靶向BPTF表达调控肺癌细胞A549增殖、迁移和侵袭

miR-454-3p regulates the proliferation,migration and invasion of A549 cancer cells by targeting BPTF

目的探讨miR-454-3p对肺癌细胞增殖、迁移和侵袭的影响及分子机制。方法采用RT-PCR技术检测miR-454-3p在肺癌组织以及肺癌细胞株中的表达,以表达量最低的肺癌细胞A549为后续分析对象,将miR-454-3p mimic转入A549细胞,RT-PCR验证miR-454-3p的过表达效率;采用CCK-8、迁移、侵袭等实验,观察转染组与对照组肺癌细胞的增殖、迁移和侵袭情况;生物信息学预测BPTF是miR-454-3p的靶标,构建BPTF 3′UTR荧光素酶载体,通过双荧光素酶报告基因验证miR-454-3p和BPTF的靶向关系;应用Western blot法检测BPTF的表达及迁移、侵袭相关蛋白的变化。结果RT-PCR实验表明miR-454-3p在肺癌组织和细胞中表达下调,且在A549细胞中表达最低(P<0.05)。与对照组相比,过表达miR-454-3p的肺癌细胞A549增殖能力明显降低,迁移和侵袭能力均受到抑制(P<0.05)。生物信息学软件分析BPTF为miR-454-3p的潜在靶基因,过表达miR-454-3p后BPTF的表达水平明显受到抑制。同时,迁移相关蛋白MMP-2和MMP-9表达明显下调,E-cadhern表达明显上调,而E-cadherin的负性调控N-cadherin表达下降。结论miR-454-3p可能通过靶向下调BPTF的表达,抑制肺癌细胞的增殖、迁移和侵袭能力,为肺癌的靶向治疗提供潜在靶标。

Purpose To investigate the effects of miR-454-3p on proliferation,migration and invasion of lung cancer cells and to explore the possible molecular mechanisms.Methods The expressions of miR-454-3p in lung cancer tissues and cells were detected by RT-PCR.The miR-454-3p mimic was transfected into A549 cells which was lowest expression in lung cancer cells.The overexpression efficiency of miR-454-3p was verified by RT-PCR.The proliferation,migration and invasion of cells in the transfection group and the control group were observed using the CCK-8,migration and invasion assay.BPTF was predicted as the target of miR-454-3p by bioinformatics.3′UTR luciferase vector of BPTF was constructed and verified the target relationship between miR-454-3p and BPTF by dual-luciferase reporter assay.The expression of BPTF was detected by Western blot.Western blot was used to detect the expression of protein involved in the cell proliferation,migration and invasion.Results RT-PCR results showed that the expression of miR-454-3p was down-regulated in lung cancer tissues and cell lines and was lowest in A549 cells.Compared with the control group,overexpression of miR-454-3p could inhibit the proliferation,migration and invasion of A549(P<0.05).BPTF was predicted as the target of miR-454-3p by bioinformatics.The expression level of BPTF was significantly inhibited after the overexpression of miR-454-3p.At the same time,the expression of MMP-2 and MMP-9 involved in the migration by miR-454-3p overexpression was also increased,and the expression of E-cadherin related with cell invasion was decreased,N-cadherin,the negative regulatory protein of E-cadherin was increased.Conclusion miR-454-3p inhibit proliferation,migration and invasion of lung cancer cells by targeting BPTF expression,providing a potential target for targeted treatment of lung cancer.