Liu Caihong, Yang Huan, Xu Zhijie, Li Dan, Zhou Meiyu, Xiao Kui, Shi Zhihui, Zhu Lanyan, Yang Lifang, Zhou Rui
Department of Respiratory Medicine, Second Xiangya Hospital, Central South University, Changsha, China.
J Cancer Res Clin Oncol. 2015 Mar;141(3):431-41. doi: 10.1007/s00432-014-1836-7. Epub 2014 Sep 23.
microRNAs (miRNAs) have been documented playing a critical role in cancer development and progression. In this study, we investigated the role of miR-548l in non-small cell lung cancer (NSCLC) migration and invasion.
microRNAs microarray analysis was used to detect the differentially expressed miRNAs between various metastatic levels of NSCLC cells and further confirmed by real-time PCR analysis. To facilitate the delineation of the role of selected miR-548l in NSCLC pathology, we detected its expression in 22 NSCLC tissues. Proliferation, apoptosis, invasion and metastasis effects of the miRNA were evaluated using MTT, flow cytometry, wound healing and invasion assay following transfection with mimics and inhibitors. Luciferase assay and Western blot analysis were performed to assess miR-548l binding to AKT1 gene. AKT1 expression in the clinical tissues was evaluated using immunohistochemical staining.
The results showed a negative relationship between miR-548l expression and lymph node metastasis of NSCLC. Functional assays showed that over-expression of miR-548l suppressed NSCLC cell migration and invasion. Luciferase assays confirmed that miR-548l could directly bind to the 3' untranslated region of AKT1. Further data showed that the over-expression of AKT1 could rescue the effects of miR-548l in NSCLC cells, and the miR-548l expression was inversely correlated with AKT1 expression in NSCLC tissues. These results indicated that AKT1 was involved in miR-548l-induced suppression of NSCLC cell migration and invasion.
These results suggested that miR-548l may play a causal role through AKT1 in NSCLC invasion and metastasis.
已有文献证明微小RNA(miRNA)在癌症发生和发展中起关键作用。在本研究中,我们调查了miR-548l在非小细胞肺癌(NSCLC)迁移和侵袭中的作用。
使用miRNA微阵列分析检测NSCLC细胞不同转移水平之间差异表达的miRNA,并通过实时PCR分析进一步确认。为了便于阐明所选miR-548l在NSCLC病理学中的作用,我们检测了其在22例NSCLC组织中的表达。在用模拟物和抑制剂转染后,使用MTT、流式细胞术、伤口愈合和侵袭试验评估miRNA的增殖、凋亡、侵袭和转移作用。进行荧光素酶测定和蛋白质印迹分析以评估miR-548l与AKT1基因的结合。使用免疫组织化学染色评估临床组织中AKT1的表达。
结果显示miR-548l表达与NSCLC的淋巴结转移呈负相关。功能测定表明,miR-548l的过表达抑制了NSCLC细胞的迁移和侵袭。荧光素酶测定证实miR-548l可直接结合AKT1的3'非翻译区。进一步的数据表明,AKT1的过表达可以挽救miR-548l对NSCLC细胞的作用,并且miR-548l表达与NSCLC组织中AKT1表达呈负相关。这些结果表明AKT1参与了miR-548l诱导的NSCLC细胞迁移和侵袭的抑制。
这些结果表明,miR-548l可能通过AKT1在NSCLC侵袭和转移中起因果作用。