miR-1273g-3p通过靶向CNR1调节A549细胞迁移能力  

MiR-1273g-3p Regulates A549 Cell Migration by Targeting CNR1

作　　者：侯绿滨[1] 苏雪婷[1] 秦性良[1] 丁红梅[1] 黄皑雪[1] 李慧[1] 葛兴枫 赵强[1] 李洁[1] 邵宁生[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京市100850

出　　处：《医学分子生物学杂志》2014年第3期125-131,共7页Journal of Medical Molecular Biology

基　　金：国家重点基础研究发展计划（973计划）（No.2011CB811300,2010CB912801）,国家自然科学基金（No.31100569,30971630,31370794）

摘　　要：目的寻找hsa-miR-1273g-3p在人肺腺癌细胞A549中的特异性靶标并探索其生物学功能。方法定量PCR（qPCR）检测miR-1273g-3p在人肺癌细胞A549、H460和H1299中表达情况，利用生物信息学方法预测miR-1273g-3p靶基因，qPCR、免疫印迹（Western Blot）和荧光素酶实验确证靶标，划痕实验和MTS实验验证miR-1273g-3p的生物学功能。结果miR-1273g-3p在非小细胞肺癌细胞系A549、I-1460和H1299中高表达：qPCR、Western印迹和荧光素酶实验证实miR-1273g-3p能够靶向CNR1，可以在mRNA与蛋白水平上调控CNR1的表达；miR-1273g-3p能够通过改变CNR1的表达水平影响A549的细胞迁移能力，对细胞增殖影响不明显。结论研究证明CNR1是miR-1273g-3p的靶基因，揭示miR-1273g-3p通过靶向CNR1影响A549细胞迁移的新功能。Objective To identify the target gene of hsa-miR-1273g-3p and investigate its biological function in human A549 cells. Methods The expression level of miR-1273g-3p in A549, H460 and H1299 cells was detected by quantitative real-time PCR （qRT-PCR） . The target gene of miR-1273g-3p was predicted by bioinformatics and confirmed by qRT-PCR, Western blotting and luciferase report assay. The wound-healing assay and MTS assay were used to examine the biological function of miR-1273g-3p. Results miR-1273g-3p was over-expressed in A549, H460 and H1299 cells, qRT-PCR, Western blotting and luciferase report assay confirmed that miR-1273g-3p could target CNR1 and it could regulate CNR1 at mRNA and protein level. MiR-1273g-3p could promote the migration of A549 ceils by inhibiting the expression of CNR1. Its inhibitory effect on the proliferation of A549 ceils was not significant. Conclusion CNR1 is a target gene of miR-1273g-3p. MiR- 1273g-3p can promote A549 cell migration by targeting CNR1.

关 键 词：miR-1273g-3p CNR1 细胞迁移 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]