Analysis of Genetic Variants and the ceRNA Network of miR-9 in Non-Small Cell Lung Cancer
Analysis of Genetic Variants and the ceRNA Network of miR-9 in Non-Small Cell Lung Cancer
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DOI：
10.1089/dna.2021.0549
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发表时间：
2022-02-01
影响因子：
3.1
通讯作者：
Wu，Jianqing
中科院分区：
文献类型：
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作者：
Wu，Shuangshuang;Xu，Jiali;Wu，Jianqing
目的：探讨hsa-miR-9单核苷酸多态性（single nucleotide polymorphism，SNPs）在非小细胞肺癌（non-small cell lung cancer，NSCLC）中的作用。方法：采用Log-rank和考克斯回归分析方法，分析miR-9基因4个功能性SNPs与中国NSCLC患者总生存期（overall survival，OS）的关系。进行报告荧光素酶测定以检查SNP与miR-9的转录活性之间的关系。实时荧光定量PCR检测细胞中miR-9的表达。在裸鼠中建立异种移植模型，用Lv-miR-9-模拟物或Lv-miR-9-抑制剂处理。结果：rs 1501672与1001例中国NSCLC患者的预后相关（A>G，相加模型：校正风险比= 0.89，95%可信区间= 0.79- 1.00，p = 0.056）。在293 T、SPC-A1和A549细胞系中，野生型A等位基因的荧光素酶活性高于突变型G等位基因。肺癌细胞中miR-9的表达水平显著高于正常肺细胞。根据癌症基因组图谱和基因表达综合数据库，miR-9在肺癌组织中也过表达。基于H1299细胞的异种移植模型显示，与lv-miR-9-NC组相比，lv-miR-9-抑制剂显著降低肿瘤生长（p< 0.001）。结论：miR-9的rs 1501672与中国人群NSCLC患者的预后相关。lncRNA-miR-9-mRNA ceRNA网络揭示了NSCLC潜在的分子生物学调控途径和预后生物标志物。
Objective:To explore the role of single-nucleotide polymorphisms (SNPs) in hsa-miR-9 in non-small cell lung cancer (NSCLC).Methods:Log-rank and Cox regression analyses were conducted to assess the association of four functional SNPs of miR-9 with overall survival (OS) of Chinese patients with NSCLC. A reporter luciferase assay was performed to examine the relationship between the SNPs and transcriptional activity of miR-9. The expression of miR-9 in cells was detected by quantitative real-time PCR assay. Xenograft model was established in nude mice, which were treated with Lv-MiR-9-mimics or Lv-miR-9-inhibitor. A long noncoding RNA (lncRNA)-miR-9-messenger RNA (mRNA) competing endogenous RNA (ceRNA) network was established based on bioinformatics analyses.Results:We found that rs1501672 was associated with the prognosis of 1001 Chinese NSCLC patients (A>G, additive model: adjusted hazard ratio = 0.89, 95% confidence interval = 0.79–1.00,p= 0.056). Luciferase reporter assay showed higher luciferase activity with wild A allele than that with mutant G allele in 293T, SPC-A1, and A549 cell lines. The miR-9 level was significantly higher in lung cancer cells than normal lung cells. miR-9 was also over expressed in lung cancer tissue according to The Cancer Genome Atlas and gene expression omnibus databases. Xenograft models based on H1299 cells showed that lv-miR-9-inhibitor significantly decreased tumor growth compared with the lv-miR-9-NC group (p< 0.001). Bioinformatics analysis showed that one target gene leukemia inhibitory factor receptor and two lncRNAs (KIAA0087andGVINP1) were associated with OS of NSCLC patients.Conclusion:The rs1501672 of miR-9 was associated with the prognosis of NSCLC patients in the Chinese population. The lncRNA-miR-9-mRNA ceRNA network revealed potential molecular biological regulation pathways and prognostic biomarkers for NSCLC.