The long non-coding RNA DKFZp434J0226 regulates the alternative splicing process through phosphorylation of SF3B6 in PDAC.
The long non-coding RNA DKFZp434J0226 regulates the alternative splicing process through phosphorylation of SF3B6 in PDAC.
复制标题
长非编码 RNA DKFZp434J0226 通过 PDAC 中 SF3B6 的磷酸化调节选择性剪接过程
DOI：
10.1186/s10020-021-00347-7
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发表时间：
2021-08-28
期刊：
影响因子：
--
通讯作者：
Xue R
中科院分区：
文献类型：
--
作者：
Li J;Tong H;Li D;Jiang Q;Zhang Y;Tang W;Jin D;Chen S;Qin X;Zhang S;Xue R
背景
长链非编码RNA（longnoncodingRNAs，lncRNA）是一类广泛存在的调控多种生物学过程的基因，在不同类型的恶性肿瘤中表达差异。lncRNA在胰腺导管腺癌（PDAC）发生中的作用尚不清楚。在此，我们研究了lncRNA DKFZp 434 J 0226在PDAC中的作用。
方法
使用微阵列分析来分析六个PDAC和配对非肿瘤组织中异常表达的mRNA和lncRNA。实时定量聚合酶链反应用于评估PDAC组织中DKFZp 434 J 0226的表达。采用CCK-8法、伤口愈合法、软琼脂集落形成法和transwell法检测PDAC细胞的侵袭力和增殖能力。此外，RNA下拉，免疫荧光，RNA免疫沉淀和蛋白质印迹分析进行了研究DKFZp 434 J 0226和SF 3B 6之间的关联。使用小鼠中的肿瘤异种移植物来测试体内肿瘤形成。
结果
在我们的研究中，222种mRNA和128种lncRNA异常表达（≥ 2倍变化）。其中，66种mRNA和53种lncRNA上调，75种lncRNA和156种mRNA下调。KEGG通路分析和Gene ontology分类表明这些基因与mRNA选择性剪接和代谢平衡的调节有关。临床分析显示，DKFZp 434 J 0226的过表达与肿瘤分级差、频繁的神经周围浸润、晚期肿瘤淋巴结转移阶段以及总生存期和进展时间缩短相关。功能测定表明DKFZp 434 J 0226在体外促进PDAC细胞迁移、侵袭和生长，并在体内加速肿瘤增殖。DKFZp 434 J 0226与剪接因子SF 3B 6相互作用，促进SF 3B 6磷酸化，进而调节前体mRNA的选择性剪接。
结论
这项研究表明，DKFZp 434 J 0226通过磷酸化SF 3B 6在PDAC中调节选择性剪接，并导致PDAC中的致癌表型。
Background
Long noncoding RNAs (lncRNAs), a type of pervasive genes that regulates various biological processes, are differentially expressed in different types of malignant tumors. The role of lncRNAs in the carcinogenesis of pancreatic ductal adenocarcinoma (PDAC) remains unclear. Here, we investigated the role of the lncRNA DKFZp434J0226 in PDAC.
Methods
Aberrantly expressed mRNAs and lncRNAs among six PDAC and paired non-tumorous tissues were profiled using microarray analysis. Quantitative real-time polymerase chain reaction was used to evaluate DKFZp434J0226 expression in PDAC tissues. CCK-8 assay, wound-healing assay, soft agar colony formation assay, and transwell assay were performed to assess the invasiveness and proliferation of PDAC cells. Furthermore, RNA pull-down, immunofluorescence, RNA immunoprecipitation, and western blotting assays were performed to investigate the association between DKFZp434J0226 and SF3B6. Tumor xenografts in mice were used to test for tumor formation in vivo.
Results
In our study, 222 mRNAs and 128 lncRNAs were aberrantly expressed (≥ twofold change). Of these, 66 mRNAs and 53 lncRNAs were upregulated, while 75 lncRNAs and 156 mRNAs were downregulated. KEGG pathway analysis and the Gene ontology category indicated that these genes were associated with the regulation of mRNA alternative splicing and metabolic balance. Clinical analyses revealed that overexpression of DKFZp434J0226 was associated with worse tumor grading, frequent perineural invasion, advanced tumor-node-metastasis stage, and decreased overall survival and time to progression. Functional assays demonstrated that DKFZp434J0226 promoted PDAC cell migration, invasion, and growth in vitro and accelerated tumor proliferation in vivo. Mechanistically, DKFZp434J0226 interacted with the splicing factor SF3B6 and promoted its phosphorylation, which further regulated the alternative splicing of pre-mRNA.
Conclusions
This study indicates that DKFZp434J0226 regulates alternative splicing through phosphorylation of SF3B6 in PDAC and leads to an oncogenic phenotype in PDAC.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
--
作者：
Liu, Jiang-Hua;Chen, Gang;Luo, Dian-Zhong
通讯作者：
Luo, Dian-Zhong
影响因子：
6.6
作者：
Siebring-van Olst E;Blijlevens M;de Menezes RX;van der Meulen-Muileman IH;Smit EF;van Beusechem VW
通讯作者：
van Beusechem VW
影响因子：
7.4
作者：
Sana J;Faltejskova P;Svoboda M;Slaby O
通讯作者：
Slaby O
DOI：
10.1158/1940-6207.capr-11-0299
发表时间：
2012-03
期刊：
Cancer prevention research (Philadelphia, Pa.)
影响因子：
--
作者：
Bao B;Wang Z;Ali S;Ahmad A;Azmi AS;Sarkar SH;Banerjee S;Kong D;Li Y;Thakur S;Sarkar FH
通讯作者：
Sarkar FH