miR-103靶向PTEN并激活PI3K/AKT通路促进肺癌细胞A549对达沙替尼耐药

miR-103 targets PTEN to promote dasatinib resistance in lung cancer A549 cells via activating PI3K/AKT pathway

目的:探讨miR-103靶向PTEN并激活PI3K/AKT信号通路促进肺癌细胞对达沙替尼(dasatinib,DASA)耐药的机制。方法:收集2014年4月至2018年1月昆明医科大学第一附属医院胸外科收治的资料完整的肺癌DASA耐药组织和不耐药组织各35例。采用q PCR实验检测miR-103在肺癌DASA耐药组织和细胞中的表达水平,同时,采用CCK-8、Transwell和Wb实验检测敲降miR-103对A549/DASA细胞增殖、迁移和上皮间质转化(EMT)的影响,双荧光素酶报告基因验证miR-103与PTEN的靶向关系。进一步采用CCK-8、Transwell和Wb实验检测miR-103通过PTEN-PI3K/AKT信号通路对A549/DASA细胞恶性生物学行为的影响。结果:miR-103在肺癌DASA耐药组织和A549/DASA细胞中均高表达(均P<0.01)。敲降miR-103可显著抑制A549/DASA细胞的增殖、迁移和EMT(P<0.05或P<0.01)。此外,双荧光素酶报告基因证实miR-103靶向作用PTEN并下调其表达水平(P<0.01)。进一步实验显示,过表达mi R-103通过靶向下调PTEN并激活PI3K/AKT信号通路进而显著促进A549/DASA细胞增殖、迁移和EMT(P<0.05或P<0.01),从而上调A549/DASA细胞对DASA的耐药性。结论:miR-103/PTEN/PI3K/AKT信号通路与肺癌DASA耐药性存在调控关系,敲降miR-103可逆转A549/DASA对DASA耐药。

Objective:To explore the mechanism of miR-103 targeting PTEN(gene of phosphate and tension homology deleted on chromsome ten)and activating PI3K/AKT signaling pathway to promote dasatinib(DASA)resistance in lung cancer cells.Methods:DASA-resistant tissues and non-resistant tissues(35 samples for each)from patients treated in Department of Thoracic Surgery,the First Affiliated Hospital of Kunming Medical University from April 2014 to January 2018 were collected for this study.Expression of miR-103 was detected in DASA-resistant tissues and cell lines of lung cancer by quantitative Real-time polymerase chain reaction(qPCR).The effect of miR-103 knock-down on the proliferation,invasion and epithelial mesenchymal transition(EMT)of A549/DASA cells were measured by CCK-8 assay,Transwell and Wb,respectively.Subsequently,the dual luciferase reporter gene assay was used to verify whether PTEN was a target gene of miR-103.CCK-8,Transwell and Wb assay were further used to investigate the effect of miR-103 on malignant biological behaviors of A549/DASA cells via regulating PTEN-PI3K/AKT signaling pathway.Results:miR-103 was highly expressed in DASA-resistant tissues and A549/DASA cells(P<0.01).Knockdown of miR-103 significantly inhibited the proliferation,invasion and EMT of A549/DASA cells(P<0.05 or P<0.01).Additionally,dual luciferase reporter gene assay confirmed that miR-103 directly targeted PTEN and down-regulated its expression(P<0.01).Mechanistically,over-expression of miR-103 targeted and down-regulated PTEN to promote cell viability,invasion and EMT via activating PI3K/AKT pathway(P<0.05 or P<0.01),and further up-regulated the DASA-resistance in A549/DASA cells.Conclusion:miR-103/PTEN/PI3K/AKT signaling pathway plays a certain role in regulating DASA resistance of lung cancer,and knockdown of miR-103 expression may reverse the resistance of A549/DASA cells to DASA.