肺腺癌组织中miR-206的表达及其对肺腺癌细胞迁移侵袭的影响

MiR-206 inhibits A549 cells migration and invasion by targeting SOX4 in lung adenocarcinoma

目的研究miR-206在人肺腺癌组织中的表达情况，探讨miR-206是否通过下调SOX4影响肺腺癌细胞系（A549）细胞的侵袭。方法收集94例肺腺癌组织标本及癌旁正常肺组织，通过实时荧光定量PCR（RT—PCR）检测各组组织中与肺腺癌相关的miR-206、miR-21、miR-34、miR-78、miR-138、miR-32、SOX4mRNA的表达情况，Pearson相关分析检验miR-206表达量与SOX4的相关性；免疫印迹法检测两组标本中SOX4的表达情况，进一步在细胞实验中，通过转染miR-206mimic和miR-NC至离体培养的A549细胞中，免疫印迹法检测A549细胞中MMP-2、MMP-9、SOX4表达情况，划痕试验和Transwell法观察细胞迁移侵袭情况，利用荧光素酶素试验验证SOX4是miR-206的靶基因；再通过转染过表达SOX4的质粒至A549细胞，检测A549细胞中MMP-2、MMP-9、SOX4的表达情况，划痕试验和Transwell法观察细胞迁移侵袭情况。结果相比于正常肺组织，miR-206、miR-21在肺腺癌组织中的表达降低（P〈0．01）；miR-34、miR-378、miR．138、miR-32的表达在两组之间没有明显区别；肺腺癌组织中SOX4mRNA的表达增加（P〈0．01）；miR-206与SOX4mRNA的表达量成负相关（r=-0．344，P〈0．01）。在细胞实验中，相比于miR—NC组，转染miR-206mimic后，A549细胞中MMP-2、MMP-9、SOX4的表达减少（P〈0．05），A549细胞的迁移侵袭减少（P〈0．01）；荧光素酶试验结果显示，miR-206能降低SOX4—3-UTR质粒的荧光素活性（P〈0．05）；转染过表达SOX4质粒后，相比于miR-206＋空质粒组，A549细胞中MMP-2和MMP-9表达增加（P〈0．05），细胞的迁移侵袭增加（P〈0．01）。结论在肺腺癌组织中，miR-206低表达；miR-206可以通过下调SOX4从而抑制A549的迁移和侵袭。

Objective To verify whether miR-206 could inhibit A549 cells migration and invasion by targeting SOX4 in lung adenocarcinoma. Methods Collection normal lung tissue and lung adenocarcinoma tissue from 94 patients. RT-PCR was used to test the expression of miR-206, miR-21, miR-34, miR-378, miR-138, miR-32 and SOX4 mRNA; Pearson correlation analysis was performed to test the correlation of miR-206 with SOX4 mRNA. In vitro experiment, after miR-206 was transfected into A549 cells, wound healing were employed to test the migratory ability of A549 cells, Transwen were used to test the invasion ability of A549 cells, Western blotting were used to investigate the expressions of MMP-2, MMP-9 and SOX4 in A549 cell. Luciferase assay was used to confirmed whether SOX4-3 -UTR the target gene of miR-206. Results Compared with normallung tissue, the expression of miR-206 and miR-21 was decreased in lung adenocarcinoma （P 〈 0.01 ）, meanwhile SOX4 mRNA was significantly increased in lung adenocarcinoma（ P 〈 0.01 ）. Pearson correlation analysis showed that miR-206 had a significantly negative correlation with SOX4 mRNA （ r = - 0. 344, P 〈 0.01 ）. In the vitro experiment, compared with the miR-206 group, the expression of MMP-2, MMP-9 and SOX4 were decreased in the miR-206 mimic group（P 〈0.01 ） ; The migration activity of A.549 cells was decreased in the miR-206 mimie group （ P 〈 0.01 ） ; The Luciferase activity of the SOX4-3 -UTR plasmid was significantly suppressed by miR-206 （P 〈 0.00） ; Over expression of SOX4 could reverse the effect of miR-206 on A549. Conclusion In lung adenocarcinoma tissue, the expression of miR-206 decreased ; miR-206 inhibited A549 cells migration and invasion by targeting SOX4-3-UTR.