敲减miR-9抑制肺非小细胞癌细胞侵袭和迁移

Knockdown of miR-9 Inhibits Invasion and Migration of Non-Small Cell Lung Cancer Cells

目的研究敲减miR-9对肺非小细胞癌(NSCLC)细胞侵袭和迁移的影响及其机制。方法收集46例NSCLC患者(包含15例T1期、15例T2期和16例T3+T4期),用RT-qPCR法检测NSCLC组织和和癌旁组织中miR-9的表达水平。用RT-qPCR法检测人正常肺上皮细胞(BEAS-2B)以及NSCLC细胞株A549和NCI-H1299中miR-9的表达水平。将miR-9-inhibitor转染入A549和NCI-H1299细胞,用CCK-8法检测细胞增殖,用Transwell法检测细胞迁移与侵袭。用生物信息法、双荧光素酶法和功能法研究miR-9的靶基因。结果与癌旁组织比较,miR-9在NSCLC组织中表达明显升高(P<0.001),且其表达随T分期递增(P<0.01)。与BEAS-2B比较,miR-9在A549和NCI-H1299细胞中表达明显升高(P<0.001)。敲减miR-9可显著降低A549和NCI-H1299细胞的增殖、侵袭和迁移(P<0.01或P<0.001)。TET2是miR-9的靶基因。转染si-TET2可逆转miR-9敲减对A549细胞迁移与侵袭的抑制作用(P<0.01)。结论敲减miR-9通过TET2抑制NSCLC细胞增殖、迁移与侵袭。

Objective To investigate the effect of miR-9 knockdown on invasion and migration of non-small cell lung cancer(NSCLC)cells and its mechanisms.Methods The expression of miR-9 in NSCLC tissues and paracancerous tissues was detected by RT-qPCR in 46 patients(15 patients with stage T1 NSCLC,15 patients with stage T2 NSCLC,and 16 patients with stage T3+T4 NSCLC).Furthermore,miR-9 expression was determined in human normal lung epithelial cell line(BEAS-2B)and NSCLC cell lines(A549 and NCI-H1299).After the miR-9-inhibitor was transfected into A549 and NCI-H1299 cells,cell proliferation was detected by CCK-8 assay and cell migration and invasion were measured by transwell assay.The target gene of miR-9 was studied by bioinformatics,double luciferase and functional assay.Results The expression of miR-9 in NSCLC tissues was higher than that in paracancerous tissues(P<0.001).Furthermore,the expression of miR-9 increased with T stage(P<0.01).Compared with BEAS-2B cells,miR-9 expression increased in both A549 and NCI-H1299 cells(P<0.001).Moreover,miR-9 knockdown reduced the proliferation,invasion and migration of A549 and NCI-H1299 cells(P<0.01 or P<0.001).TET2 was the target gene of miR-9.Transfection with si-TET2 reversed the inhibitory effect of miR-9 knockdown on the migration and invasion of A549 cells(P<0.01).ConclusionKnockdown of miR-9 inhibits the proliferation,migration and invasion of NSCLC cells through TET2.