目的：探究microRNA-101 (miR-101)通过靶向成纤维细胞生长因子2（fibroblast growth factor 2，FGF2）抑制非小细胞肺癌（non-small cell lung cancer，NSCLC）细胞迁移和侵袭的分子机制。方法：采用qPCR法检测人正常肺上皮细胞BEAS-2B和NSCLC细胞系A549、H661和SK-MES-1，以及转染后A549细胞miR-101和FGF2的表达水平。分别将miR-NC、miR-101 mimics、miR-IN-NC、miR-101 inhibitor或pcDNA-3.1空质粒、pcDNA-FGF2转染至A549细胞，运用划痕愈合实验和Transwell小室实验，检测A549细胞中过表达miR-101和FGF2对NSCLC细胞迁移和侵袭能力的影响，采用Western blotting（WB）法检测各组A549细胞中FGF2、E-cadherin、N-cadherin、Vimentin、ERK1/2和p-ERK1/2的表达水平。结果：miR-101在NSCLC细胞系中的表达水平明显低于正常肺上皮细胞（均P<0.05），而以A549细胞中表达水平为最低。过表达miR-101可明显抑制A549细胞的迁移（P<0.05）和侵袭（P<0.01），且使细胞中E-cadherin的表达增多（P<0.05）而Vimentin（P<0.05）、N-cadherin（P<0.01）和p-ERK1/2（P<0.05）的表达水平降低。抑制miR-101表达后，可以显著增强A549细胞的迁移和侵袭能力（均P<0.05），引起细胞中E-cadherin表达明显降低而 Vimentin、N-cadherin 和 p-ERK1/2 表达水平增高（均 P<0.05）。采用 WB 法和双荧光素酶报告基因实验验证了 FGF2 是miR-101的直接靶基因，且过表达FGF2后显著增强A549细胞的迁移和侵袭能力（均P<0.01），以及减少细胞中E-cadherin的表达（P<0.01）而增加Vimentin（P<0.01）、N-cadherin（P<0.05）和p-ERK1/2（P<0.05）表达水平。与单独过表达FGF2组相比，共同过表达miR-101和FGF2组A549细胞迁移和侵袭能力明显减弱（均P<0.01），其E-cadherin的表达增多（P<0.01）而Vimentin（P<0.01）、N-cadherin（P<0.05）和p-ERK1/2表达水平下降（P<0.01）。结论：miR-101通过调控靶基因FGF2抑制NSCLC A549细胞的上皮间质转化（EMT）过程及ERK信号通路，进而抑制NSCLC细胞的迁移和侵袭。

Objective: To investigate the molecular mechanism of microRNA-101 (miR-101) inhibiting the migration and invasion of non-small cell lung cancer (NSCLC) via targeting fibroblast growth factor 2 (FGF2). Methods: qPCR was used to detect the expression levels of miR-101 and FGF2 in human normal lung epithelial BEAS-2B cells and NSCLC cell lines (A549, H661 and SK-MES-1) as well as A549 cells after transfection. MiR-NC, miR-101 mimics, miR-IN-NC, miR-101 inhibitor or pcDNA-3.1 empty plasmid,pcDNA-FGF2 were respectively transfected into A549 cells. Wound healing assay and Transwell assay were used to examine the effects of overexpression of miR-101 and FGF2 on the migration and invasion of A549 cells. Western blotting(WB) was used to detect the expression levels of FGF2, E-cadherin, N-cadherin, Vimentin, ERK1/2 and p-ERK1/2 in A549 cells in each group. Results: The expression level of miR-101 in NSCLC cell lines were significantly lower than that in normal lung epithelial cells (all P<0.05), while the expression level in A549 cells was the lowest. Overexpression of miR-101 significantly inhibited the migration (P<0.05) and invasion (P<0.01) of A549 cells, increased the expression level of E-cadherin but decreased the expression level of Vimentin (P<0.05),N-cadherin (P<0.01) and p-ERK1/2 (P<0.05). Inhibition of miR-101 significantly enhanced the invasion and migration of A549 cells (all P<0.05), decreased the expression level of E-cadherin but increased the expression levels of Vimentin, N-cadherin and p-ERK1/2 (all P<0.05). The results of WB and Dual-luciferase reporter gene assay verified that FGF2 is a direct target gene of miR-101, and over‐expression of FGF2 significantly enhanced the invasion and migration of A549 cells (all P<0.01), decreased the expression of E-cad‐herin (P<0.01) but increased the expressions of Vimentin (P<0.01), N-cadherin (P<0.05) and p-ERK1/2 (P<0.01). Compared with the FGF2 overexpression alone group, co-overexpression of miR-101 and FGF2 significantly reduced the invasion and migration of A549 cells (all P<0.01), increased the expression of E-cadherin (P<0.01), and decreased the expressions of Vimentin (P<0.01), N-cadherin (P<0.05) and p-ERK1/2 (P<0.01). Conclusion: By targeting FGF2, miR-101 inhibits the invasion and migration of NSCLC cells through suppressing the epithelial-mesenchymal transition (EMT) and ERK signaling pathway.

国家自然科学基金资助项目（No.81372519）；教育部博士点基金资助项目（No.20131202110005）；天津市自然科学基金重点资助项目（No.18JCZDJC98500）