Gao Yang, Yang Fan
General Practice Center, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu 610000, China
General Practice Center, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu 610000, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2017 Jul 28;42(7):755-761. doi: 10.11817/j.issn.1672-7347.2017.07.003.
To investigate the effect of miR-26b on the invasion and migration of lung cancer cell and to explore its mechanism. Methods: qPCR was used to detect the expression of miR-26b in lung cancer. Luciferase reporter gene was used to detect interaction between miR-26b and hENT1. Transwell assay was used to detect invasion ability after treatment of miR-26b mimics. Scratch assay was used to detect migration ability after treatment of miR-26b mimics. The expressions of hENT1, ROCK-1 and RhoA were detected by Western blot. The changes of cytoskeleton after miR-26b mimics treatment with phalloidin were observed. The effect of miR-26b mimics on the tumor size and volume of lung cancer was determined by subcutaneous tumor formation in nude mice. Results: MiR-26b expression was significantly reduced in lung cancer. With the progress of lung cancer, the expression of miR-26b was reduced. With the progress in differentiation of lung cancer, the expression of miR-26b was decreased. Decrease of miR-26b was associated with lung cancer lymph node metastasis. HENT1 was the direct target of miR-26b; miR-26b regulated the invasion and migration ability of human lung carcinoma A549 cells. MiR-26b regulated the expression of hENT1, ROCK-1 and RhoA. After the treatment with miR-26b mimics, the F-actin staining was significantly reduced, whereas the formation of wrinkles and the formation of pseudopodia were significantly reduced. Subcutaneous tumor formation in nude mice showed that miR-26b mimics treatment significantly reduced the tumor size and mass. Conclusion: MiR-26b plays a role in tumor suppression in lung cancer. miR-26b can regulate the invasion and migration ability of lung carcinoma A549 cells by targeting hENT1 depending on the RhoA/ROCK-1 pathway.
探讨miR-26b对肺癌细胞侵袭和迁移的影响并探究其机制。方法：采用qPCR检测肺癌中miR-26b的表达。运用荧光素酶报告基因检测miR-26b与hENT1之间的相互作用。采用Transwell实验检测miR-26b模拟物处理后的侵袭能力。采用划痕实验检测miR-26b模拟物处理后的迁移能力。通过蛋白质免疫印迹法检测hENT1、ROCK-1和RhoA的表达。用鬼笔环肽观察miR-26b模拟物处理后细胞骨架的变化。通过裸鼠皮下成瘤实验确定miR-26b模拟物对肺癌肿瘤大小和体积的影响。结果：肺癌中miR-26b表达显著降低。随着肺癌进展，miR-26b表达降低。随着肺癌分化程度进展，miR-26b表达下降。miR-26b降低与肺癌淋巴结转移相关。hENT1是miR-26b的直接靶点；miR-26b调节人肺癌A549细胞的侵袭和迁移能力。miR-26b调节hENT1、ROCK-1和RhoA的表达。用miR-26b模拟物处理后，F-肌动蛋白染色显著减少，而皱纹形成和伪足形成显著减少。裸鼠皮下成瘤实验表明，miR-26b模拟物处理显著减小肿瘤大小和质量。结论：miR-26b在肺癌中发挥肿瘤抑制作用。miR-26b可通过RhoA/ROCK-1途径靶向hENT1调节肺癌A549细胞的侵袭和迁移能力。
