LINC00319低表达通过靶向调控miR-129-5p对非小细胞肺癌A549细胞放射敏感性的影响

The Effect of LINC00319 on the Radiosensitivity of Non- small Cell Lung Cancer A549 Cells by Targeting the Regulation of miR-129-5p

目的探讨LINC00319低表达靶向调控miR-129-5p对非小细胞肺癌A549细胞放射敏感性的影响。方法采用实时荧光定量PCR检测人永生化肺上皮细胞株16HBE和非小细胞肺癌细胞株A549中LINC00319、miR-129-5p的表达水平及LINC00319对A549细胞中miR-129-5p表达的影响,采用双荧光素酶报告基因实验验证LINC00319和miR-129-5p的靶向关系,采用克隆形成实验检测LINC00319低表达或miR-129-5p过表达对A549细胞放射敏感性的影响;给予4Gy X射线照射A549细胞后,采用流式细胞术检测LINC00319低表达或miR-129-5p过表达对A549细胞凋亡的影响。结果与人永生化肺上皮细胞株16HBE相比,非小细胞肺癌细胞株A549中LINC00319表达水平明显升高,而miR-129-5p的表达水平明显降低(P<0.05);LINC00319过表达可使A549细胞中miR-129-5p表达降低,而LINC00319低表达则使miR-129-5p表达升高;双荧光素酶报告基因实验证实miR-129-5p是LINC00319的潜在靶基因;LINC00319低表达或者miR-129-5p过表达可降低A549细胞的存活分数,并促进4GyX射线照射诱导的A549细胞凋亡。结论 LINC00319低表达可通过靶向调控miR-129-5p增强非小细胞肺癌A549细胞放射敏感性。

Objective To investigate the effect of LINC00319 on the radiosensitivity of non-small cell lung cancer A549 cells by targeting the regulation the expression of miR-129-5 p.Methods Real-time quantitative PCR was applied to detect the expression levels of LINC00319 and miR-129-5 p in human immortalized lung epithelial cell line 16 HBE and non-small cell lung cancer cell line A549, and the effect of LINC00319 on the expression of miR-129-5 p in A549 cells. The relationship between LINC00319 and miR-129-5 p was validated by double luciferase reporter gene assay.The effects of LINC00319 low expression or miR-129-5 p overexpression on the radiosensitivity of A549 cells were determined by cloning experiment. Specifically, after 4 Gy X-ray irradiation to A549 cells, flow cytometry was performed to evaluate the effects of LINC00319 low expression or miR-129-5 p overexpression on apoptosis of A549 cells.Results Compared with human immortalized pulmonary epithelial cell line 16 HBE, the expression of LINC00319 in non-small cell lung cancer cell line A549 was significantly increased, while the expression of miR-129-5 p was greatly decreased(P<0.05).Over-expression of LINC00319 could reduce the expression of miR-129-5 p in A549 cells, while low expression of LINC00319 could improve the expression of miR-129-5 p in A549 cells. Luciferase reporter gene assay confirmed that miR-129-5 p was a potential targeted gene of LINC00319. LINC00319 low expression or miR-129-5 p overexpression could reduce the survival rate of A549 cells and promote apoptosis of A549 cells induced by 4 Gy X-ray irradiation.Conclusion Low expression of LINC00319 can enhance the radiosensitivity of non-small cell lung cancer A549 cells through targeting the regulation of miR-129-5 p.