Jiang Min, Zhong Ting, Zhang Wei, Xiao Zuke, Hu Guozhu, Zhou Hong, Kuang Haibin
Department of Respiration, Jiangxi Province People's Hospital, Nanchang, Jiangxi 330006, P.R. China.
Department of Physiology, School of Medicine, Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Mol Med Rep. 2017 May;15(5):3231-3238. doi: 10.3892/mmr.2017.6398. Epub 2017 Mar 28.
Previous studies have demonstrated that microRNA (miR)-205-5p expression is significantly increased in non‑small cell lung cancer tissues and is associated with tumor differentiation grade. The aim of the present study was to explore the effects of miR‑205‑5p on viability, apoptosis and invasion of lung cancer A549 cells. The hsa‑miR‑205‑5p small interfering RNA (siRNA) inhibitor was transfected into A549 cells and expression of miR‑205‑5p was detected by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Cell viability, apoptosis and invasion were assayed by Cell Counting kit‑8, Annexin V/propidium iodide double staining and Transwell assay, respectively. Target genes of miR‑205‑5p were predicted using bioinformatics analysis. Expression of mRNA and protein levels of candidate target genes following miR‑205‑5p inhibition were detected using RT‑qPCR and western blot analysis respectively. The results demonstrated that relative survival rates of A549 cells were significantly inhibited in miR‑205‑5p siRNA‑transfected cells at 24 and 48 h compared with control cells. Apoptosis was markedly increased in the miR‑205‑5p siRNA cells compared with control cells. The number of invaded cells following miR‑205‑5p siRNA silencing was significantly decreased compared with control cells. Bioinformatics analysis revealed that erb‑B2 receptor kinase 3 (erbB3), zinc finger E‑box binding homeobox 2 (ZEB2), clathrin heavy chain (CLTC) and mediator complex subunit 1 (MED1) may be potential target genes of miR‑205‑5p. Reduced expression of miR‑205‑5p significantly increased the expression of ZEB2 mRNA and protein, inhibited the expression of erbB3 protein, but had no significant effect on the expression levels of CLTC and MED1. In summary, reduced expression of miR‑205‑5p promoted apoptosis and inhibited proliferation and invasion in lung cancer A549 cells through upregulation of ZEB2 and downregulation of erbB3. The present results suggested that the increased miR‑205‑5p expression observed in non‑small cell lung cancer tissues may contribute to increased proliferation and invasion of lung cancer cells and thus to cancer progression.
先前的研究表明，微小RNA（miR）-205-5p在非小细胞肺癌组织中的表达显著增加，且与肿瘤分化程度相关。本研究的目的是探讨miR-205-5p对肺癌A549细胞活力、凋亡和侵袭的影响。将hsa-miR-205-5p小干扰RNA（siRNA）抑制剂转染至A549细胞中，并通过逆转录-定量聚合酶链反应（RT-qPCR）检测miR-205-5p的表达。分别通过细胞计数试剂盒-8、膜联蛋白V/碘化丙啶双染法和Transwell实验检测细胞活力、凋亡和侵袭情况。使用生物信息学分析预测miR-205-5p的靶基因。在抑制miR-205-5p后，分别使用RT-qPCR和蛋白质印迹分析检测候选靶基因的mRNA和蛋白质水平的表达。结果表明，与对照细胞相比，在转染miR-205-5p siRNA的细胞中，A549细胞在24小时和48小时的相对存活率显著受到抑制。与对照细胞相比，miR-205-5p siRNA细胞中的凋亡明显增加。与对照细胞相比，miR-205-5p siRNA沉默后侵袭细胞的数量显著减少。生物信息学分析显示，表皮生长因子受体-2（erbB3）、锌指E盒结合同源框蛋白2（ZEB2）、网格蛋白重链（CLTC）和中介体复合物亚基1（MED1）可能是miR-205-5p的潜在靶基因。miR-205-5p表达的降低显著增加了ZEB2 mRNA和蛋白质的表达，抑制了erbB3蛋白质的表达，但对CLTC和MED1的表达水平没有显著影响。总之，miR-205-5p表达的降低通过上调ZEB2和下调erbB3促进了肺癌A549细胞的凋亡，并抑制了其增殖和侵袭。目前的结果表明，在非小细胞肺癌组织中观察到的miR-205-5p表达增加可能有助于肺癌细胞增殖和侵袭的增加，从而促进癌症进展。
