Identification of miRNAs that specifically target tumor suppressive KLF6-FL rather than oncogenic KLF6-SV1 isoform
Identification of miRNAs that specifically target tumor suppressive KLF6-FL rather than oncogenic KLF6-SV1 isoform
复制标题
鉴定特异性靶向肿瘤抑制性 KLF6-FL 而不是致癌 KLF6-SV1 亚型的 miRNA。
DOI：
10.4161/rna.29356
复制
发表时间：
2014-07-01
期刊：
影响因子：
4.1
通讯作者：
Waye, Mary Miu-Yee
中科院分区：
文献类型：
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作者：
Liang, Wei-Cheng;Wang, Yan;Waye, Mary Miu-Yee
Kruppel like factor6(KLF6)基因编码多种不同的蛋白质亚型，其中大部分与肝癌的发生和肿瘤的发展密切相关。最近的生物信息学分析表明，KLF6基因的mRNA选择性剪接产生了大约16个具有不同甚至相反功能的选择性剪接变体。有趣的是，全长KLF6(KLF6-FL)是一种在肝癌中经常失活的抑癌基因，而KLF6剪接变异体(KLF6-SV1)是一种具有拮抗KLF6-FL功能的癌基因亚型。令人信服的证据表明，miRNA是一种小的内源性非编码RNA(NcRNA)，通过靶向蛋白质编码基因的不同mRNA区域，在调节多种细胞生物学过程中发挥重要作用。为了确定潜在的针对KLF6-FL的miRNAs，我们利用生物信息学分析结合荧光素酶报告基因的分析，筛选出两个miRNAs，即miR-210和miR-1301，它们专门针对肿瘤抑制的KLF6-FL而不是致癌的KLF6-SV1。我们的体外实验表明，稳定表达KLF6-FL抑制细胞的增殖、迁移和血管生成，而过表达miR-1301促进细胞迁移和血管生成。进一步的实验证明miR-1301在肝癌细胞系和临床标本中都有高表达，我们还鉴定了miR-1301基因的潜在甲基化和组蛋白乙酰化。综上所述，我们的发现揭示了一种新的分子机制，即特定的miRNAs通过靶向肿瘤抑制亚型KLF6-FL而不是其致癌亚型KLF6-SV1来促进肿瘤的发生。
The Kruppel like factor 6 (KLF6) gene encodes multiple protein isoforms derived from alternative mRNA splicing, most of which are intimately involved in hepatocarcinogenesis and tumor progression. Recent bioinformatics analysis shows that alternative mRNA splicing of the KLF6 gene produces around 16 alternatively spliced variants with divergent or even opposing functions. Intriguingly, the full-length KLF6 (KLF6-FL) is a tumor suppressor gene frequently inactivated in liver cancer, whereas KLF6 splice variant 1 (KLF6-SV1) is an oncogenic isoform with antagonistic function against KLF6-FL. Compelling evidence indicates that miRNA, the small endogenous non-coding RNA (ncRNA), acts as a vital player in modulating a variety of cellular biological processes through targeting different mRNA regions of protein-coding genes. To identify the potential miRNAs specifically targeting KLF6-FL, we utilized bioinformatics analysis in combination with the luciferase reporter assays and screened out two miRNAs, namely miR-210 and miR-1301, specifically targeted the tumor suppressive KLF6-FL rather than the oncogenic KLF6-SV1. Our in vitro experiments demonstrated that stable expression of KLF6-FL inhibited cell proliferation, migration and angiogenesis while overexpression of miR-1301 promoted cell migration and angiogenesis. Further experiments demonstrated that miR-1301 was highly expressed in liver cancer cell lines as well as clinical specimens and we also identified the potential methylation and histone acetylation for miR-1301 gene. To sum up, our findings unveiled a novel molecular mechanism that specific miRNAs promoted tumorigenesis by targeting the tumor suppressive isoform KLF6-FL rather than its oncogenic isoform KLF6-SV1.