摘要
背景与目的已有的研究表明miR-424可抑制肾癌细胞增殖,抑制宫颈鳞癌细胞的迁移和侵袭能力,而其对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞的影响目前尚无系统研究。本研究探讨miR-424对NSCLC A549细胞生长和侵袭迁移能力的影响并进一步研讨其分子机制。方法应用CCK8检测过表达及抑制miR-424的表达对A549细胞增殖的影响。应用Transwell检测过表达及抑制miR-424的表达对A549细胞侵袭能力的影响。应用Western blot检测过表达及抑制miR-424的表达对A549细胞中MMP9和MMP2蛋白水平的影响。构建E2F6 3’UTR区的荧光素酶报告载体,验证miR-424对E2F6的靶向作用。采用Western blot检测过表达及抑制miR-424的表达后,A549细胞中E2F6的表达。结果过表达miR-424后,A549的生长和侵袭能力显著降低。过表达miR-424后,A549细胞的MMP-2和MMP-9表达下降。荧光素酶活性检测表明miR-424能够抑制E2F6的荧光素酶活性。过表达miR-424后,细胞内E2F6的表达降低。结论 miR-424能够通过调控E2F6而抑制A549的生长和侵袭能力。
Background and objectivehTe inhibitory ability of miR-424 on the proliferation of renal carcinoma cell and the migration and invasion of cancer cells has been widely explored and demonstrated. However, the effects of miR-424 on non-small cell lung cancer （NSCLC） have not been systematically examined. In this study, detected the growth and inva-sion effect of miR-424 in NSCLC A549 cell. hTe migration and molecular mechanism of this cell are also detected.Methods NSCLC A549 cell was transfected with miR-424 and its inhibitor. Atfer transfection, the proliferation ability of A549 cell was detectedby CCK8 assay. hTen, the migration ability in A549 cell was detected by migration assays. Furthermore, the expression level of MMP2 and MMP9 in A549 was detected by Western blot and immune lfuorescence. hTe 3＆#39;UTR of E2F6 was cloned into luciferase reporter vector and its enzymatic activitywas detected to verify whether miR-424 can target E2F6. hTe expres-sion level of E2F6 in a549 cell atfer transfecing with miR-424 was detected by Western blot.Results Atfer transfection of miR-424, the proliferation and migration abilities were remarkably decreased and the expression level of MMP-2 and MMP-9 were down-regulated in A549. Moreover, MiR-424 inhibited the enzymatic activity of luviferase reporter vector of E2F6. Spe-ciifcally, the expression level of E2F6 was down-regulated in A549.ConclusionmiR-424 can inhibit the proliferation and migration abilities of A549 by negatively regulating the expression of E2F6.
出处
《中国肺癌杂志》
CAS
CSCD
北大核心
2016年第9期571-576,共6页
Chinese Journal of Lung Cancer
