miR-520a-3p通过靶向FZD8增强非小细胞肺癌A549/TAX细胞对紫杉醇的敏感性  

miR-520a-3p enhances the paclitaxel sensitivity of non-small cell lung cancer A549/TAX cells by targeting FZD8

作　　者：禹玺[1] 李晓平[2] 钱雪娇[1] 刘江波[1] 郑洪[1] YU Xi;LI Xiaoping;QIAN Xuejiao;LIU Jiangbo;ZHENG Hong(Department of Respiratory, the First Central Hospital of Tianjin City,Tianjin 300092,China;Department of Thoracic Surgery,the First Central Hospital of Tianjin City,Tianjin 300092,China)

机构地区：[1]天津市第一中心医院呼吸科,天津300092 [2]天津市第一中心医院胸外科,天津300092

出　　处：《中国肿瘤生物治疗杂志》2019年第10期1107-1112,共6页Chinese Journal of Cancer Biotherapy

摘　　要：目的:探讨miR-520a-3p通过靶向卷曲受体8(FZD8)调控非小细胞肺癌(NSCLC)A549/TAX细胞对紫杉醇(TAX)的敏感性。方法:选用NSCLC A549细胞、TAX抗性细胞株A549/TAX及人肺上皮细胞HLF-α,用qPCR检测A549和A549/TAX细胞中miR-520a-3p的表达水平。依据转染质粒的不同分为对照组、miR-520a-3p mimics组、si-FZD8组和si-FZD8+miR-520a-3p inhibitor组,用6μmol/L的TAX处理各组A549/TAX细胞后,用CCK-8检测A549/TAX细胞的增殖能力,用Annexin V-FITC/PI染色流式细胞术检测A549/TAX细胞的凋亡水平,用WB检测A549/TAX细胞中FZD8的表达水平。双荧光素酶报告基因系统检测miR-520a-3p与FZD8的靶向关系。结果:miR-520a-3p在A549/TAX细胞中低表达(P<0.01),且TAX能够上调A549/TAX细胞中miR-520a-3p的表达水平(P<0.01)。6μmol/L TAX处理后,过表达miR-520a-3p可显著抑制A549/TAX细胞的增殖并促进其凋亡(均P<0.01)。双荧光素酶报告基因证明miR-520a-3p可靶向下调FZD8的表达水平(P<0.01)。si-FZD8可显著抑制A549/TAX细胞增殖、促进细胞凋亡,从而增强细胞对TAX的敏感性;同时敲降miR-520a-3p和FZD8可逆转敲降FZD8对A549/TAX细胞TAX敏感性的增强作用(P<0.01)。结论:miR-520a-3p可通过靶向下调FZD8表达水平增强A549/TAX细胞TAX敏感性。Objective:To investigate the influence of miR-520a-3p on paclitaxel(TAX)sensitivity of non-small cell lung cancer(NSCLC)A549/TAX cells via regulating frizzled class receptor 8(FZD8).Methods:NSCLC A549 cells,TAX-resistant cell line A549/TAX and human lung epithelial HLF-αcells were selected.The expression level of miR-520a-3p in A549 and A549/TAX cells was detected by qPCR.According to different transfection plasmids,the experimental cells were divided into control group,miR-520a-3p mimics group,si-FZ8 group and si-FZD8+miR-520a-3p inhibitor group.After being treated with 6μmol/L paclitaxel,the proliferation of A549/TAX cells was determined by CCK-8 assay.Flow cytometry with Annexin V-FLTC/PI staining was used to detect the apoptosis level of A549/TAX cells.The expression of FZD8 in A549/TAX cells was detected by WB.The targeting relationship between miR-520a-3p and FZD8 was verified by the dual-luciferase reporter gene system.Results:miR-520a-3p was poorly expressed in TAX-resistant A549/TAX cells(P<0.01),and TAX up-regulated the expression of miR-520a-3p in A549/TAX cells(P<0.01).After the treatment with 6μmol/L TAX,over-expression of miR-520a-3p significantly inhibited the proliferation of A549/TAX cells and promoted apoptosis(all P<0.01).Dual luciferase reporter gene assay showed that miR-520a-3p targetedly down-regulated the expression of FZD8(P<0.01).si-FZD8 could significantly inhibit the proliferation and promote cell apoptosis of A549/TAX cells,thereby enhancing the TAX sensitivity of cells.At the same time,simultaneous knockdown of miR-520a-3p and FZD8 could reverse the enhancement of FZD8 knockdown on TAX sensitivity of A549/TAX cells(P<0.01).Conclusion:miR-520a-3p enhances the TAX sensitivity of A549/TAX cells by down-regulating the expression of FZD8.

关 键 词：非小细胞肺癌 紫杉醇 敏感性 miR-520a-3p 卷曲受体8 

分 类 号：R734.2[医药卫生—肿瘤] R730.5[医药卫生—临床医学]