MicroRNA-1258 suppresses tumour progression via GRB2/Ras/Erk pathway in non-small-cell lung cancer
MicroRNA-1258 suppresses tumour progression via GRB2/Ras/Erk pathway in non-small-cell lung cancer
复制标题
MicroRNA-1258 通过 GRB2/Ras/Erk 通路抑制非小细胞肺癌的肿瘤进展。
DOI：
10.1111/cpr.12502
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发表时间：
2018-12-01
影响因子：
8.5
通讯作者：
Chen, Yijiang
中科院分区：
文献类型：
--
作者：
Jiang, Wei;Wei, Ke;Chen, Yijiang
目的肺癌在世界范围内仍是一种高发病率和高死亡率的疾病。MicroRNA已被证明在多种实体瘤的再利用中起着不可或缺的作用。尽管miR-1258在抑制乳腺癌和胃癌转移中起着重要作用，但miR-1258在非小细胞肺癌中的具体生物学功能尚不清楚。方法采用qRT-PCR和原位杂交技术检测miR-1258在NSCLC组织及癌旁组织中的差异表达。进行流式细胞术和CCK-8、EdU、小管形成和衰老测定，并研究异种移植模型以探索miR-1258的功能。通过双荧光素酶报告基因测定、qRT-PCR、IHC和Western印迹验证miR-1258的潜在靶点。结果体外和体内功能获得和丧失实验表明，miR-1258抑制NSCLC细胞增殖，诱导细胞衰老和凋亡。荧光素酶报告基因检测、免疫组化和Western blotting分析表明，GRB 2是miR-1258的直接靶点之一。GRB 2过表达质粒可逆转miR-1258过表达后的功能变化。相反，miR-1258抑制剂显著逆转si-GRB 2诱导的GRB 2下调。从机制上讲，miR-1258的过表达抑制GRB 2表达，然后导致Ras/Erk致癌途径的失活。结论miR-1258可通过靶向GRB 2/Ras/Erk通路抑制NSCLC的进展，这可能会为NSCLC患者的潜在生物标志物和新的治疗策略提供不同的见解。
Objectives Lung cancer is still a disease with high morbidity and mortality in the world. MicroRNAs have been proven to act as an indispensable role in the reuse of multiple solid tumours. Although miR-1258 plays a vital role in suppressing metastasis in breast cancer and gastric cancer, the specific biological function of miR-1258 in non-small-cell lung cancer remains unclear. Methods The differential expression of miR-1258 in NSCLC tissues and corresponding paracancerous tissues was detected by qRT-PCR and ISH. Flow cytometry and CCK-8, EdU, tubule formation, and senescence assays were performed, and xenograft models were studied to explore the function of miR-1258. Potential targets of miR-1258 were verified by dual luciferase reporter assay, qRT-PCR, IHC and Western blotting. Results In vitro and in vivo gain- and loss-of-function assays suggested that miR-1258 inhibits NSCLC cell proliferation and induces senescence and apoptosis. The luciferase reporter assay, IHC and Western blotting analysis showed that GRB2 is one of the direct targets of miR-1258. The GRB2 overexpression plasmid can reverse the functional changes after overexpression of miR-1258. In contrast, miR-1258 inhibitor significantly reversed si-GRB2-induced GRB2 down-regulation. Mechanistically, overexpression of miR-1258 inhibits GRB2 expression and then leads to inactivation of the Ras/Erk oncogenic pathway. Conclusions Our results indicate that miR-1258 can suppress NSCLC progression by targeting the GRB2/Ras/Erk pathway, which may lead to different insights into potential biomarkers and novel therapeutic strategies for NSCLC patients.