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[miR-758-3p通过靶向NUSAP1抑制非小细胞肺癌细胞的增殖和侵袭]

[miR-758-3p inhibits the proliferation and invasion of non-small cell lung cancer cells by targeting NUSAP1].

作者信息

Li X M, Yu Z, Huai M, Han H Y, Cao S S

机构信息

Department of Pathology, North China Petroleum Administration General Hospital, Renqiu 062552, China.

Department of Cardiothoracic, North China Petroleum Administration General Hospital, Renqiu 062552, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2021 Jan 23;43(1):113-117. doi: 10.3760/cma.j.cn112152-20190727-00472.

Abstract

To investigate the biological function of miR-758-3p and the underlying mechanism in non-small cell lung cancer (NSCLC). miR-758-3p mimics was transfected to A549 NSCLC cells, miRNA control was used as a negative control, cells transfected with nucleolar and spindle associated protein 1 (NUSAP1)-overexpression vector indicated as NUSAP1 group, cells co-transfected with miR-758-3p mimics and NUSAP1-overexpression vector indicated as miR-758-3p mimics+ NUSAP1 group. The effects of miR-758-3p on the proliferation, migration and invasion abilities of A549 cells were detected by cell counting kit-8 (CCK-8) and Transwell assays, respectively. Bioinformatics and dual luciferase reporter assay were used to predict and verify the target gene of miR-758-3p. The expressions of miR-758-3p and NUSAP1 in A549 cells were significantly up-regulated at 24 hours after transfection with miR-758-3p mimics (<0.05). Compared with the miRNA control group (1.15±0.06), the OD value of miR-758-3p mimic group (0.78±0.06) was significantly decreased at 72 hours after transfection (<0.05). The number of migrated cells of miR-758-3p mimic group (119.04±11.49) was significantly lower than that of the control group (271.38±19.05) (<0.05). The number of invaded cells of miR-758-3p mimic group (71.33±5.36) was significantly lower than the control group (164.30±8.11) (<0.05). The result of dual-luciferase reporter assay showed that NUSAP1 was a direct target of miR-758-3p. Moreover, up-regulation of NUSAP1 abolished the inhibitory effects of miR-758-3p on the proliferation, migration and invasion abilities of A549 cells. miR-758-3p inhibits the proliferation, migration and invasion of NSCLC cells by targeting NUSAP1.

摘要

探讨miR-758-3p在非小细胞肺癌(NSCLC)中的生物学功能及潜在机制。将miR-758-3p模拟物转染至A549非小细胞肺癌细胞,以miRNA对照作为阴性对照,转染核仁与纺锤体相关蛋白1(NUSAP1)过表达载体的细胞作为NUSAP1组,共转染miR-758-3p模拟物和NUSAP1过表达载体的细胞作为miR-758-3p模拟物+NUSAP1组。分别采用细胞计数试剂盒-8(CCK-8)和Transwell实验检测miR-758-3p对A549细胞增殖、迁移和侵袭能力的影响。运用生物信息学和双荧光素酶报告基因实验预测并验证miR-758-3p的靶基因。转染miR-758-3p模拟物24小时后,A549细胞中miR-758-3p和NUSAP1的表达显著上调(<0.05)。转染72小时后,与miRNA对照组(1.15±0.06)相比,miR-758-3p模拟物组的光密度值(OD值)(0.78±0.06)显著降低(<0.05)。miR-758-3p模拟物组的迁移细胞数(119.04±11.49)显著低于对照组(271.38±19.05)(<0.05)。miR-758-3p模拟物组的侵袭细胞数(71.33±5.36)显著低于对照组(164.30±8.11)(<0.05)。双荧光素酶报告基因实验结果表明,NUSAP1是miR-758-3p的直接靶基因。此外,NUSAP1的上调消除了miR-758-3p对A549细胞增殖、迁移和侵袭能力的抑制作用。miR-758-3p通过靶向NUSAP1抑制非小细胞肺癌细胞的增殖、迁移和侵袭。

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