miR-140-5p靶向HDAC7抑制非小细胞肺癌细胞增殖、迁移和侵袭的机制探讨

目的:探讨miR-140-5p靶向组蛋白去乙酰化酶7(histone deacetylase 7,HDAC7)调控非小细胞肺癌（non-small cell lung cancer,NSCLC）细胞增殖、迁移和侵袭的机制。方法:采用qRT-PCR检测人NSCLC组织及癌旁组织中miR-140-5p的表达水平。用miR-140-5p mimic（模拟物）及miR-140-5p NC（阴性对照）转染A549细胞，CCK8法及克隆形成实验检测细胞增殖能力，划痕实验和Transwell实验检测细胞迁移和侵袭能力,双荧光素酶报告基因实验验证miR-140-5p与HDAC7的靶向关系，Western blot检测各组细胞HDAC7及PI3K、p-PI3K、AKT、和p-AKT表达水平。结果:与癌旁组织相比，miR-140-5p在NSCLC组织中的表达水平明显减低，差异有统计学意义（P＜0.01）。与miR-140-5p NC组相比，过表达miR-140-5p后A549细胞在48和72 h的增殖能力明显降低（P＜0.05）；且细胞克隆形成、细胞迁移和侵袭能力均降低（均P＜0.05），PI3K/AKT信号通路中关键分子p-PI3K和p-AKT蛋白水平明显降低（均P＜0.05)。生物信息学预测HDAC7可能是miR-140-5p的一个靶基因，且双荧光素酶报告结果证实miR-140-5p直接靶向调节HDAC7表达。结论:miR-140-5p通过靶向HDAC7表达进而抑制NSCLC A549细胞的增殖、迁移和侵袭，其机制可能与通过抑制PI3K/AKT信号通路的激活有关。

Mechanism of miR-140-5p targeting HDAC7 inhibiting proliferation,migration and invasion of non-small cell lung cancer cells

Objective:To explore the mechanism of miR-140-5p targeting HDAC7 regulating proliferation,migration and invasion of non-small cell lung cancer cells.Methods:The expression level of miR-140-5p in human NSCLC tissues and adjacent tissues was detected by qRT-PCR.A549 cells were transfected with miR-140-5p mimic（mimic) and miR-140-5p NC（negative control).Cell proliferation was detected by CCK8 assay and colony formation assay.Cell migration and invasion were detected by scratch test and Transwell test.The double luciferase target experiment verified the targeting relationship between miR-140-5p and HDAC7.Western blot was used to detect the expression levels of HDAC7,PI3K,p-PI3K,AKT and p-AKT.Results:The expression level of miR-140-5p in NSCLC tissues was significantly lower than that in adjacent tissues（P<0.01).Compared with miR-140-5p NC group,the proliferation ability of A549 cells at 48 and 72 h after overexpression of miR-140-5p significantly decreased（P<0.05),the ability of cell clone formation,cell migration and invasion were decreased（all P<0.05),and the protein levels of p-PI3K and p-AKT in PI3K/AKT signaling pathway were significantly decreased（all P<0.05).Bioinformatics predicted that HDAC7 might be a target gene of miR-140-5p,and the results of double luciferase assay confirmed that miR-140-5p could directly target the expression of HDAC7.Conclusion:miR-140-5p inhibits the proliferation,migration and invasion of NSCLC A549 cells by targeting HDAC7 expression,which may be related to the inhibition of PI3K/AKT signaling pathway.