Chen Ping, Zhao Yunlong, Li Yingjie
First Affiliated Hospital of PLA General Hospital, Beijing 100048, China.
Zhongguo Fei Ai Za Zhi. 2017 Jul 20;20(7):452-458. doi: 10.3779/j.issn.1009-3419.2017.07.03.
To explore the function and the potential molecular mechanism of miR-218 in lung cancer cell.
The expression of miR-218 mRNA was determined by real-time PCR in lung cancer tissues, adjacent tissues and lung cancer cells. Transwell assay was used to detect the migration and invasion of A549 cell after transfected with Anti-miR-218 or negative control and HC4006 cell after transfected with miR-218 mimics and miR-218 negative control. Targetscan and MiRanda were used to calculate the potential targets of miR-218 and Luciferase reporter assay was performed to identify that the Robo1 was one target genes of miR-218. Transwell assay was used to detect whether miR-218 regulated the invasion of lung cancer cell transfected with anti-miR-218 or negative control via Robo1.
The expression of miR-218 in the lung cancer tissues was significantly lower than that in the adjacent tissues (P<0.05). Inhibition of miR-218 improved the migration and invasion of A549 cell. Overexpression of miR-218 suppressed the migration and invasion of HCC4006 cell. The co-transfection of anti-miR-218 or miR-218 mimics and the Robo1 3'UTR increased or reduced the luciferase activity of Robo1 compared with the control group (P<0.05). Inhibition of miR-218 and Robo1 recovered the invaded cells of A549. Overexpression of miR-218 and inhibition of Robo1 reduced the number of the invased cells of HCC4006. These results suggested that miR-218 banded Robo1 directly and inhibited lung cancer cell invasion by targeting Robo1.
CONCLUSIONS: MiR-218 inhibited the migration and invasion of lung cancer cells through regulating Robo1 expression. .
探讨miR-218在肺癌细胞中的作用及其潜在分子机制。
采用实时定量PCR检测肺癌组织、癌旁组织及肺癌细胞中miR-218 mRNA的表达。运用Transwell实验检测转染Anti-miR-218或阴性对照后的A549细胞以及转染miR-218模拟物和miR-218阴性对照后的HC4006细胞的迁移和侵袭能力。利用Targetscan和MiRanda软件预测miR-218的潜在靶标,并通过荧光素酶报告基因实验鉴定Robo1是miR-218的靶基因之一。采用Transwell实验检测miR-218是否通过Robo1调控转染anti-miR-218或阴性对照的肺癌细胞的侵袭能力。
肺癌组织中miR-218的表达显著低于癌旁组织(P<0.05)。抑制miR-218可促进A549细胞的迁移和侵袭。过表达miR-218可抑制HCC4006细胞的迁移和侵袭。与对照组相比,共转染anti-miR-218或miR-218模拟物与Robo1 3'UTR可增加或降低Robo1的荧光素酶活性(P<0.05)。抑制miR-218和Robo1可使A549细胞的侵袭细胞数量恢复。过表达miR-218并抑制Robo1可减少HCC4006细胞的侵袭细胞数量。这些结果表明miR-218直接结合Robo1并通过靶向Robo1抑制肺癌细胞侵袭。
miR-218通过调控Robo1表达抑制肺癌细胞的迁移和侵袭。