miR-758-3p通过靶向核仁纺锤体相关蛋白1抑制非小细胞肺癌细胞增殖和侵袭

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目的:探讨miR-758-3p在非小细胞肺癌(NSCLC)中的生物学功能及其作用机制。方法:采用脂质体法转染miR-758-3p模拟物(miR-758-3p模拟物组)、miRNA对照(对照组)、核仁纺锤体相关蛋白1(NUSAP1)过表达质粒(NUSAP1过表达组)、miR-758-3p模拟物+NUSAP1(miR-758-3p模拟物+NUSAP1组)至人NSCLC细胞系A549细胞中。采用CCK-8法和Transwell小室法检测A549细胞增殖、迁移和侵袭能力,结合生物信息学预测网站和双荧光素酶报告基因实验验证miR-758-3p及其靶基因的靶向关系。结果:转染24 h后,miR-758-3p模拟物组和对照组A549细胞中miR-758-3p基因的相对表达水平分别为3.02±0.16和1.00±0.08,NUSAP1基因的相对表达水平分别为0.04±0.02和1.00±0.03,差异均有统计学意义(均n P<0.05)。转染72 h后,miR-758-3p模拟物组和对照组细胞的吸光度值分别为0.78±0.06和1.15±0.06,差异有统计学意义(n P<0.05)。miR-758-3p模拟物组细胞的迁移数和侵袭数分别为[(119.04±11.49)个]和[(71.33±5.36)个],均低于对照组[分别为(271.38±19.05)个和(164.30±8.11)个;均n P<0.05]。miR-758-3p模拟物转染野生型NUSAP1报告基因的荧光素酶活性(0.37±0.04)低于对照组(1.00±0.03,n P0.05)。n 结论:miR-758-3p通过调控NUSAP1基因的表达抑制NSCLC细胞的增殖、迁移和侵袭。“,”Objective:To investigate the biological function of miR-758-3p and the underlying mechanism in non-small cell lung cancer (NSCLC).Methods: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.Results: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 (n P<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 (n P<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) (n P<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) (n P<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.n Conclusion:miR-758-3p inhibits the proliferation, migration and invasion of NSCLC cells by targeting NUSAP1.
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