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微小RNA-183/-96/-182簇在大多数乳腺癌中上调，并增加细胞增殖和迁移。

MiR-183/-96/-182 cluster is up-regulated in most breast cancers and increases cell proliferation and migration.

作者信息

Li Pei, Sheng Cheng, Huang Lingling, Zhang Hui, Huang Lihua, Cheng Zeneng, Zhu Qubo

出版信息

Breast Cancer Res. 2014 Nov 14;16(6):473. doi: 10.1186/s13058-014-0473-z.

DOI:10.1186/s13058-014-0473-z
PMID:25394902
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4303194/
Abstract

INTRODUCTION

The miR-183/-96/-182 cluster is a conserved polycistronic microRNA (miRNA) cluster which is highly expressed in most breast cancers. Although there are some sporadic reports which demonstrate the importance of each miRNA in this cluster in breast cancer, the biological roles of this cluster as a whole and its regulation mechanisms in breast cancer are still unclear. We compared the expression of this cluster in different cancer types, analyzed the regulation mechanism of this cluster, identified new target genes, and examined the impact of this cluster on breast cancer cells.

METHODS

The miRNA level was detected by LNA-based northern blot and Real-time PCR, and was also analyzed from TCGA dataset. Bioinformatics research and luciferase assay were applied to find the promoter regions and transcription factors. To investigate the biological effects of the miR-183/-96 /-182 cluster in breast cancer, we generated miR-96, miR-182 and miR-183 overexpression stable cell lines to check the overdose effects; we also used miR-Down™ antagomir for each miRNA as well as miR-183/-96 /-182 cluster sponge lentivirus to check the knockdown effects. Growth, migration, cell cycle profile and survival of these cells was then monitored by colony formation assay, MTT assay, cell wound healing assay, flow cytometry and microscopy. The target gene was validated by Real-time PCR, luciferase assay, Western blot and Phalloidin/DAPI counterstaining.

RESULTS

The miR-183/-96/-182 cluster was highly expressed in most breast cancers, and its transcription is disordered in breast cancer. The miR-183/-96/-182 cluster was transcribed in the same pri-miRNA and its transcription was regulated by ZEB1 and HSF2. It increased breast cell growth by promoting more rapid completion of mitosis, promoted cell migration and was essential for cell survival. MiR-183 targeted the RAB21 mRNA directly in breast cancer.

CONCLUSION

The miR-183/-96/-182 cluster is up-regulated in most breast cancer. It functions as an oncogene in breast cancer as it increases cell proliferation and migration.

摘要

引言

miR-183/-96/-182簇是一个保守的多顺反子微小RNA（miRNA）簇，在大多数乳腺癌中高度表达。尽管有一些零星报道表明该簇中每个miRNA在乳腺癌中的重要性，但该簇作为一个整体在乳腺癌中的生物学作用及其调控机制仍不清楚。我们比较了该簇在不同癌症类型中的表达，分析了该簇的调控机制，鉴定了新的靶基因，并研究了该簇对乳腺癌细胞的影响。

方法

通过基于锁核酸（LNA）的Northern印迹和实时定量PCR检测miRNA水平，并从癌症基因组图谱（TCGA）数据集进行分析。应用生物信息学研究和荧光素酶测定来寻找启动子区域和转录因子。为了研究miR-183/-96 /-182簇在乳腺癌中的生物学效应，我们构建了miR-96、miR-182和miR-183过表达稳定细胞系以检测过量表达的影响；我们还使用针对每个miRNA的miR-Down™反义寡核苷酸以及miR-183/-96 /-182簇海绵慢病毒来检测敲低效应。然后通过集落形成试验、MTT试验、细胞划痕愈合试验、流式细胞术和显微镜观察来监测这些细胞的生长、迁移、细胞周期分布和存活情况。通过实时定量PCR、荧光素酶测定、蛋白质免疫印迹和鬼笔环肽/4',6-二脒基-2-苯基吲哚（DAPI）复染来验证靶基因。

结果

miR-183/-96/-182簇在大多数乳腺癌中高度表达，其转录在乳腺癌中紊乱。miR-183/-96/-182簇由同一个初级miRNA转录，其转录受锌指E盒结合蛋白1（ZEB1）和热休克因子2（HSF2）调控。它通过促进有丝分裂更快完成来增加乳腺细胞生长，促进细胞迁移，并且对细胞存活至关重要。在乳腺癌中，miR-183直接靶向RAB21信使核糖核酸（mRNA）。

结论

miR-183/-96/-182簇在大多数乳腺癌中上调。它在乳腺癌中作为癌基因发挥作用，因为它增加细胞增殖和迁移。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/2d07394ba8ab/13058_2014_473_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/22373965f30a/13058_2014_473_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/28bad7a297d0/13058_2014_473_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/02b6dac81694/13058_2014_473_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/2d07394ba8ab/13058_2014_473_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/22373965f30a/13058_2014_473_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/f9d0fd32b898/13058_2014_473_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/068b910fdc78/13058_2014_473_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/e5152b0b411b/13058_2014_473_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/28bad7a297d0/13058_2014_473_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/02b6dac81694/13058_2014_473_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1da3/4303194/2d07394ba8ab/13058_2014_473_Fig7_HTML.jpg

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