miR-17-5p通过调控PKD2对肺腺癌HCC827细胞生物学行为的影响

The effect of miR-17-5p on biological behavior of lung adenocarcinoma HCC827 cells by regulating PKD2

目的:探讨miR-17-5p在肺腺癌细胞中的表达及miR-17-5p对肺腺癌细胞增殖、侵袭、迁移、凋亡的影响及其分子作用机制。方法:采用实时荧光定量PCR检测人肺腺癌细胞系HCC827和人正常肺上皮细胞BEAS-2B中miR-17-5p的表达水平。实验分为miR-17-5p过表达组(miR-17-5p mimics组)、miR-17-5p低表达组(miR-17-5p inhibitor组)和阴性对照组(NC组)。通过CCK-8、流式细胞术、Transwell实验探讨miR-17-5p对肺腺癌细胞增殖、凋亡、侵袭及迁移的影响。最后通过Targetscan数据库预测出PKD2是miR-17-5p下游潜在的靶基因,利用实时荧光定量PCR和Western-blot实验验证miR-17-5p与PKD2之间的相互作用关系。结果:miR-17-5p在HCC827细胞中高表达(P<0.001),上调miR-17-5p后显著促进了肺腺癌细胞增殖(P<0.05)、侵袭(P<0.01)及迁移(P<0.05)并抑制凋亡(P<0.01)。miR-17-5p与PKD2的3’UTR存在直接结合位点,并且过表达miR-17-5p在mRNA和蛋白水平均显著促进了PKD2的表达(P<0.01)。结论:PKD2是miR-17-5p的直接靶基因,miR-17-5p通过调控PKD2影响肺腺癌细胞的生物学行为。

Objective:To investigate the expression of miR-17-5p in lung adenocarcinoma cells,and to study the effects of miR-17-5p on proliferation,invasion,migration and apoptosis of lung adenocarcinoma cells and its molecular mechanism.Methods:The expression of miR-17-5p in human lung adenocarcinoma cell line HCC827 and human normal lung epithelial cell line BEAS-2B were detected by real-time fluorescent quantitative PCR.There were miR-17-5p mimics group,miR-17-5p inhibitor group and NC group in the experiment.The effects of miR-17-5p on proliferation,apoptosis,invasion and migration of lung adenocarcinoma cells were analyzed by CCK-8,flow cytometry and Transwell assay.Finally,PKD2 was predicted to be a potential downstream target gene of miR-17-5p by Targetscan database.The interaction between miR-17-5p and PKD2 was verified by real-time fluorescence quantitative PCR and Western-blot experiments.Results:miR-17-5p was highly expressed in HCC827 cells(P<0.001),and up-regulation of miR-17-5p significantly promoted the proliferation(P<0.05),invasion(P<0.01)and migration(P<0.05)of lung adenocarcinoma cells,and inhibited apoptosis(P<0.01).There was a direct binding site between miR-17-5p and 3'UTR of PKD2,and overexpression of miR-17-5p significantly promoted the expression of PKD2 at both mRNA and protein levels(P<0.01).Conclusion:PKD2 was the direct target gene of miR-17-5p,and miR-17-5p affected the biological behavior of lung adenocarcinoma cells by regulating PKD2.