目的：探讨长链非编码 RNA（long non-coding RNA, lncRNA）GAS6-AS2 对肺癌 A549 细胞增殖、迁移、侵袭及紫杉醇（paclitaxel，PTX）敏感性的影响及其分子机制。方法：用 qPCR 法检测肺癌 A549 和 A549/PTX 细胞中 GAS6-AS2 和miR-125a-3p的表达水平。用脂质体转染技术，分别将si-NC、si-GAS6-AS2、miR-NC、miR-125a-3p、pcDNA、pcDNA-GAS6-AS2、si-GAS6-AS2+anti-miR-NC、si-GAS6-AS2+anti-miR-125a-3p 等转染入 A549/PTX 细胞，同时用不同浓度（1、5、10、20、40 nmol/L）PTX 处理 A549 和 A549/PTX 细胞，WB 法检测细胞中增殖、迁移与侵袭相关蛋白 cyclin D1、p21、MMP2 和 MMP9 的表达水平，MTT法检测PTX对A549/PTX细胞的增殖抑制作用，Transwell 小室法检测细胞的迁移和侵袭能力。用双荧光素酶报告基因实验验证 GAS6-AS2 和 miR-125a-3p 之间的调控关系。结果: GAS6-AS2 在 A549/PTX 细胞中表达水平显著高于 A549细胞（P<0.01），miR-125a-3p在A549/PTX细胞中表达水平显著低于A549细胞（P<0.01）。不同浓度PTX处理后A549/PTX细胞的增殖抑制率显著低于A549细胞（均P<0.01），且呈浓度依赖性。抑制 GAS6-AS2 或过表达 miR-125a-3p 联合 PTX 处理均可抑制 A549/PTX 细胞的迁移和侵袭能力，增强 PTX 对细胞的增殖抑制，上调p21表达量，下调cyclin D1、MMP2和MMP9表达量（均P<0.01）。GAS6-AS2靶向负调控miR-125a-3p的表达。干扰miR-125a-3p可逆转抑制GAS6-AS2对A549/PTX细胞增殖、迁移、侵袭和PTX敏感性的作用（均P<0.01）。结论：抑制GAS6-AS2通过靶向miR-125a-3p抑制肺癌A549/PTX细胞的增殖、迁移和侵袭，增强细胞PTX敏感性，GAS6-AS2是肺癌潜在的分子靶点。

Objective: To investigate the effects and molecular mechanism of lncRNA GAS6-AS2 on the proliferation, migration and invasion of lung cancer A549 cells and its sensitivity to paclitaxel (PTX). Methods: qPCR was used to detect the levels of GAS6-AS2 and miR-125a-3p in lung cancer A549 and A549/PTX cells. Si-NC, si-GAS6-AS2, miR-NC, miR-125A-3p, pcDNA,PCDNA-GAS6-AS2, si-GAS6-AS2+anti-miR-GAS6-AS2 and si-GAS6-AS2+anti-miR-125A-3p were transfected into A549/PTX cells by liposomal transfection.A549 and A549/PTX cells were treated with PTX of different concentrations (1 nmol/L, 5 nmol/L, 10 nmol/L,20 nmol/L, 40 nmol/L). WB was used to detect the expression levels of proliferation, migration and invasion related proteins (cyclin D1, p21, MMP2 and MMP9). MTT assay was used to determine the inhibitory effect of PTX on A549/PTX cell proliferation. Transwell assay was applied to detect cell migration and invasion ability of cells. Dual-luciferase reporter gene system was performed to verify the relationship between GAS6-AS2 and miR-125a-3p. Results: The expression level of GAS6-AS2 in A549/PTX cells was significantly higher than that in A549 cells (P<0.01), and the expression level of miR-125a-3p in A549/PTX cells was significantly lower than that in A549 cells (P<0.01). The inhibitory rates of PTX at different concentrations on A549/PTX cells were significantly lower than that on A549 cells (P<0.01), in a concentration-dependent manner. GAS6-AS2 knockdown or miR-125a-3p over-expression combined with PTX treatment could inhibit A549/PTX cell migration and invasion, enhance inhibition rate of PTX on cell proliferation, promote the expression of p21 protein, and suppress the expressions of cyclin D1, MMP2 and MMP9 (all P<0.01).GAS6-AS2 targeted and negatively regulated the expression of miR-125a-3p. Interfering miR-125a-3p reversed the effects of GAS6-AS2 knockdown on proliferation, migration, invasion and PTX sensitivity of A549/PTX cells (all P<0.01). Conclusion:GAS6-AS2 knockdown inhibits proliferation, migration and invasion of A549/PTX cells and enhances sensitivity of cells to PTX by targeting miR-125a-3p; thus, it can be used as a potential molecular target for lung cancer.

海南省卫健委科研资助项目（No. 1605320273A2001）