Small RNAs targeting transcription start site induce heparanase silencing through interference with transcription initiation in human cancer cells.
Small RNAs targeting transcription start site induce heparanase silencing through interference with transcription initiation in human cancer cells.
复制标题
靶向转录起始位点的小 RNA 通过干扰人类癌细胞中的转录起始来诱导乙酰肝素酶沉默
DOI：
10.1371/journal.pone.0031379
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Tong Q
中科院分区：
文献类型：
--
作者：
Jiang G;Zheng L;Pu J;Mei H;Zhao J;Huang K;Zeng F;Tong Q
乙酰肝素酶（HPA）是一种内切-h-D-葡萄糖醛酸酶，其切割硫酸乙酰肝素蛋白聚糖的硫酸乙酰肝素链，在大多数人类癌症中过表达。最近的证据表明，小干扰RNA（siRNA）诱导人类细胞中的转录基因沉默（TGS）。在本研究中，针对位于翻译起始位点上游101 bp处的转录起始位点（TSS）的−9/+10 bp（siH 3）而非−174/−155 bp（siH 1）或−134/−115 bp（siH 2）区域转染siRNA，以序列特异性方式导致人前列腺癌、膀胱癌和胃癌细胞中乙酰肝素酶的TGS。甲基化特异性PCR和亚硫酸氢盐测序显示siH 3转染细胞中乙酰肝素酶启动子内的CpG岛没有DNA甲基化。乙酰肝素酶的TGS不涉及表观遗传标记组蛋白H3赖氨酸9二甲基化（H3 K9 me 2）、组蛋白H3赖氨酸27三甲基化（H3 K27 me 3）或活性染色质标记乙酰化组蛋白H3（AcH 3）的变化。siH 3介导的TGS不涉及选择性剪接的调节。相反，siH 3通过降低RNA聚合酶II和转录因子II B（TFIIB）的结合而干扰转录起始，但不干扰乙酰肝素酶启动子上转录因子Sp1或早期生长反应1的结合。此外，Argonaute 1和Argonaute 2促进RNA聚合酶II和TFIIB在乙酰肝素酶启动子上的结合减少，并且在siH 3诱导的乙酰肝素酶TGS中是必需的。将靶向乙酰肝素酶TSS（-9/+10 bp）的短发夹RNA构建体稳定转染到癌细胞中，导致体外和无胸腺小鼠模型中癌细胞的增殖、侵袭、转移和血管生成减少。这些结果表明，靶向TSS的小RNA可以通过干扰乙酰肝素酶的转录起始而诱导乙酰肝素酶的TGS，从而显著抑制肿瘤细胞的生长、侵袭、转移和血管生成。
Heparanase (HPA), an endo-h-D-glucuronidase that cleaves the heparan sulfate chain of heparan sulfate proteoglycans, is overexpressed in majority of human cancers. Recent evidence suggests that small interfering RNA (siRNA) induces transcriptional gene silencing (TGS) in human cells. In this study, transfection of siRNA against −9/+10 bp (siH3), but not −174/−155 bp (siH1) or −134/−115 bp (siH2) region relative to transcription start site (TSS) locating at 101 bp upstream of the translation start site, resulted in TGS of heparanase in human prostate cancer, bladder cancer, and gastric cancer cells in a sequence-specific manner. Methylation-specific PCR and bisulfite sequencing revealed no DNA methylation of CpG islands within heparanase promoter in siH3-transfected cells. The TGS of heparanase did not involve changes of epigenetic markers histone H3 lysine 9 dimethylation (H3K9me2), histone H3 lysine 27 trimethylation (H3K27me3) or active chromatin marker acetylated histone H3 (AcH3). The regulation of alternative splicing was not involved in siH3-mediated TGS. Instead, siH3 interfered with transcription initiation via decreasing the binding of both RNA polymerase II and transcription factor II B (TFIIB), but not the binding of transcription factors Sp1 or early growth response 1, on the heparanase promoter. Moreover, Argonaute 1 and Argonaute 2 facilitated the decreased binding of RNA polymerase II and TFIIB on heparanase promoter, and were necessary in siH3-induced TGS of heparanase. Stable transfection of the short hairpin RNA construct targeting heparanase TSS (−9/+10 bp) into cancer cells, resulted in decreased proliferation, invasion, metastasis and angiogenesis of cancer cells in vitro and in athymic mice models. These results suggest that small RNAs targeting TSS can induce TGS of heparanase via interference with transcription initiation, and significantly suppress the tumor growth, invasion, metastasis and angiogenesis of cancer cells.
登录
查看更多内容
影响因子：
10.8
作者：
Huang, Dan;Yang, Chongzhe;Huang, Kai
通讯作者：
Huang, Kai
影响因子：
64.5
作者：
Chen, HT;Hahn, S
通讯作者：
Hahn, S
影响因子：
14.9
作者：
Hawkins PG;Santoso S;Adams C;Anest V;Morris KV
通讯作者：
Morris KV
影响因子：
11.4
作者：
MATZKE, MA;PRIMIG, M;MATZKE, AJM
通讯作者：
MATZKE, AJM
影响因子：
16.8
作者：
Janowski, Bethany A.;Huffman, Kenneth E.;Corey, David R.
通讯作者：
Corey, David R.