Song Lei, Li Dan, Gu Yue, Wen Zhong-Mei, Jie Jing, Zhao Dan, Peng Li-Ping
Department of Respiratory Medicine, First Hospital of Jilin University, Changchun, People's Republic of China.
Department of Respiratory Medicine, First Hospital of Jilin University, Changchun, People's Republic of China.
Clin Lung Cancer. 2016 Sep;17(5):e65-e75. doi: 10.1016/j.cllc.2016.03.012. Epub 2016 Apr 6.
Our study explored whether the microRNA-126 (miR-126)-mediated PTEN/PI3K/AKT (phosphatase and tensin homology deleted on chromosome 10/phosphatidylinositol 3-kinase regulatory subunit-β/AKT) signaling pathway by targeting PIK3R2 affects the proliferation, migration, and invasion of non-small-cell lung cancer (NSCLC) A549 cells.
Quantitative real-time polymerase chain reaction was used to measure the expression of miR-126 in A549 cells. The MTT (methyl thiazolyl tetrazolium) assay, cell scratch test, Transwell assay, and Western blot were used to detect the proliferation, migration, and invasion of A549 cells and protein expression in A549 cells, respectively.
The expression of miR-126 decreased and the expression of PIK3R2 increased in A549 cells (P < .05, for both). Upregulation of miR-126 resulted in the decrease of the proliferation, migration, and invasive abilities of A549 cells, the downregulation of the expression of PIK3R2, PI3K, and phosphorylated Akt (p-Akt) protein, and the upregulation of PTEN expression (P < .05 for all). Also, these abilities of A549 cells increased, and the expression of these 3 proteins was upregulated with downregulation of miR-126 (P < .05 for all). The results of the dual luciferase reporter gene assay showed that PIK3R2 was the target gene of miR-126. PIK3R2, PI3K, and p-Akt proteins were downregulated, but PTEN protein was upregulated as PIK3R2 was silenced or the inhibitor of the PTEN/PI3K/AKT signaling pathway increased. Also, downregulation of miR-126 with silencing of PIK3R2 or increasing the inhibitor of the pathway caused increased PI3K and p-Akt protein expression and increased active proliferation, migration, and invasive abilities of A549 cells (P < .05 for all).
The upregulation of miR-126 in NSCLC A549 cells can reduce the expression of the target gene PIK3R2 and influence the PTEN/PI3K/AKT signaling pathway, suppressing the proliferation, migration, and invasive abilities of A549 cells.
我们的研究探讨了微小RNA-126（miR-126）通过靶向PIK3R2介导的PTEN/PI3K/AKT（第10号染色体上缺失的磷酸酶和张力蛋白同源物/磷脂酰肌醇3-激酶调节亚基-β/AKT）信号通路是否影响非小细胞肺癌（NSCLC）A549细胞的增殖、迁移和侵袭。
采用定量实时聚合酶链反应检测A549细胞中miR-126的表达。分别采用MTT（噻唑蓝）法、细胞划痕试验、Transwell试验和蛋白质免疫印迹法检测A549细胞的增殖、迁移和侵袭以及A549细胞中的蛋白质表达。
A549细胞中miR-126表达降低，PIK3R2表达升高（两者P均<0.05）。miR-126的上调导致A549细胞的增殖、迁移和侵袭能力降低，PIK3R2、PI3K和磷酸化Akt（p-Akt）蛋白的表达下调，PTEN表达上调（所有P均<0.05）。此外，随着miR-126的下调，A549细胞的这些能力增强，这3种蛋白的表达上调（所有P均<0.05）。双荧光素酶报告基因检测结果显示PIK3R2是miR-126的靶基因。当PIK3R2沉默或PTEN/PI3K/AKT信号通路抑制剂增加时，PIK3R2、PI3K和p-Akt蛋白下调，但PTEN蛋白上调。此外，通过沉默PIK3R2或增加该通路抑制剂下调miR-126导致PI3K和p-Akt蛋白表达增加，A549细胞的活性增殖、迁移和侵袭能力增强（所有P均<0.05）。
NSCLC A549细胞中miR-126的上调可降低靶基因PIK3R2的表达并影响PTEN/PI3K/AKT信号通路，抑制A549细胞的增殖、迁移和侵袭能力。
