The Expression of miR-375 Is Associated with Carcinogenesis in Three Subtypes of Lung Cancer.
The Expression of miR-375 Is Associated with Carcinogenesis in Three Subtypes of Lung Cancer.
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miR-375的表达与三种亚型肺癌的癌变相关
DOI：
10.1371/journal.pone.0144187
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Lu S
中科院分区：
文献类型：
--
作者：
Jin Y;Liu Y;Zhang J;Huang W;Jiang H;Hou Y;Xu C;Zhai C;Gao X;Wang S;Wu Y;Zhu H;Lu S
许多研究表明肺癌中存在独特的microRNA谱。然而，miR-375在肺癌中的作用和相关信号通路在很大程度上是未知的。我们的研究调查了腺癌、鳞状细胞癌和小细胞肺癌中癌发生与miR-375之间的关系，以确定新的治疗分子靶点。我们使用微阵列评估了来自126个速冻肺标本的显微切割癌细胞和相邻正常细胞中的723个microRNA。我们使用定量逆转录酶PCR验证了miR-375及其22个推定靶mRNA在78个速冻肺癌组织的独立队列中的表达谱。此外，我们对小细胞肺癌细胞株NCI-H82中的6个靶基因（FZD 8、ITGA 10、ITPKB、LRP 5、PIAS 1和RUNX 1）进行了双荧光素酶报告基因检测和Western blot。我们还检测了miR-375对NCI-H82细胞增殖的影响。我们发现miR-375在腺癌和小细胞肺癌中表达显著上调，而在鳞癌中表达下调。在22个推定的靶基因中，11个在3个成对比较（腺癌与正常、鳞状细胞癌与正常或小细胞肺癌与正常）中的至少2个中显示出显著不同的表达水平。6个靶基因与小细胞肺癌中miR-375的表达水平呈强负相关。进一步研究发现，miR-375直接靶向ITPKB mRNA的3 'UTR，过表达miR-375可显著降低ITPKB蛋白水平，促进细胞生长。因此，我们的研究证实了miR-375在3种肺癌亚型中的差异表达谱，并发现miR-375直接靶向ITPKB并促进SCLC细胞系的细胞生长。
Many studies demonstrated unique microRNA profiles in lung cancer. Nonetheless, the role and related signal pathways of miR-375 in lung cancer are largely unknown. Our study investigated relationships between carcinogenesis and miR-375 in adenocarcinoma, squamous cell carcinoma and small cell lung carcinoma to identify new molecular targets for treatment. We evaluated 723 microRNAs in microdissected cancerous cells and adjacent normal cells from 126 snap-frozen lung specimens using microarrays. We validated the expression profiles of miR-375 and its 22 putative target mRNAs in an independent cohort of 78 snap-frozen lung cancer tissues using quantitative reverse-transcriptase PCR. Moreover, we performed dual luciferase reporter assay and Western blot on 6 targeted genes (FZD8, ITGA10, ITPKB, LRP5, PIAS1 andRUNX1) in small cell lung carcinoma cell line NCI-H82. We also detected the effect of miR-375 on cell proliferation in NCI-H82. We found that miR-375 expression was significantly up-regulated in adenocarcinoma and small cell lung carcinoma but down-regulated in squamous cell carcinoma. Among the 22 putative target genes, 11 showed significantly different expression levels in at least 2 of 3 pair-wise comparisons (adenocarcinoma vs. normal, squamous cell carcinoma vs. normal or small cell lung carcinoma vs. normal). Six targeted genes had strong negative correlation with the expression level of miR-375 in small cell lung carcinoma. Further investigation revealed that miR-375 directly targeted the 3’UTR of ITPKB mRNA and over-expression of miR-375 led to significantly decreased ITPKB protein level and promoted cell growth. Thus, our study demonstrates the differential expression profiles of miR-375 in 3 subtypes of lung carcinomas and finds thatmiR-375 directly targets ITPKB and promoted cell growth in SCLC cell line.