Porphyromonas gingivalis Induction of MicroRNA-203 Expression Controls Suppressor of Cytokine Signaling 3 in Gingival Epithelial Cells
Porphyromonas gingivalis Induction of MicroRNA-203 Expression Controls Suppressor of Cytokine Signaling 3 in Gingival Epithelial Cells
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DOI：
10.1128/iai.00082-11
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发表时间：
2011-07-01
影响因子：
3.1
通讯作者：
Lamont, Richard J.
中科院分区：
文献类型：
--
作者：
Moffatt, Catherine E.;Lamont, Richard J.
牙龈卟啉单胞菌是严重牙周病的病原菌。能够利用原代牙龈上皮细胞(GECs)内的细胞内生活方式，牙龈假单胞菌的储存库可以在牙龈上皮细胞内持续存在。这一过程是通过操纵宿主细胞信号转导级联反应来促进的，这些信号转导通路可以影响细胞周期、细胞死亡和细胞因子反应。利用基因芯片技术，我们研究了牙龈假单胞菌33277在牙周血管内皮细胞中调节微RNA表达的能力。在牙龈假单胞菌存在的情况下，受GEC差异调控的几个miRNAs之一是miRNA-203(miR-203)，与未感染的对照组相比上调了4倍。定量逆转录-聚合酶链式反应(qRT-PCR)证实miR-203基因的差异表达。用qRT-PCR方法检测细胞因子信号转导抑制因子3(SOCS3)和SOCS6的可能靶点miR-203。与对照组相比，感染牙龈假单胞菌的GEC中SOCS3和SOCS6的mRNA水平分别降低了5倍和2倍。使用小干扰RNA构建的miR-203沉默逆转了SOCS3表达的抑制。双荧光素酶检测证实miR-203与SOCS33‘非翻译区的靶点结合。Western印迹分析表明，在miR-203沉默后，SOCS下游靶标信号转导和转录激活子3(STAT3)的激活程度降低。这项研究表明，牙龈假单胞菌诱导miRNAs可以调节重要的宿主信号反应。
Porphyromonas gingivalis is a pathogen in severe periodontal disease. Able to exploit an intracellular lifestyle within primary gingival epithelial cells (GECs), a reservoir of P. gingivalis can persist within the gingival epithelia. This process is facilitated by manipulation of the host cell signal transduction cascades which can impact cell cycle, cell death, and cytokine responses. Using microarrays, we investigated the ability of P. gingivalis 33277 to regulate microRNA (miRNA) expression in GECs. One of several miRNAs differentially regulated by GECs in the presence of P. gingivalis was miRNA-203 (miR-203), which was upregulated 4-fold compared to uninfected controls. Differential regulation of miR-203 was confirmed by quantitative reverse transcription-PCR (qRT-PCR). Putative targets of miR-203, suppressor of cytokine signaling 3 (SOCS3) and SOCS6, were evaluated by qRT-PCR. SOCS3 and SOCS6 mRNA levels were reduced > 5-fold and > 2-fold, respectively, in P. gingivalis-infected GECs compared to controls. Silencing of miR-203 using a small interfering RNA construct reversed the inhibition of SOCS3 expression. A dual luciferase assay confirmed binding of miR-203 to the putative target binding site of the SOCS3 3' untranslated region. Western blot analysis demonstrated that activation of signal transducer and activator of transcription 3 (Stat3), a downstream target of SOCS, was diminished following miR-203 silencing. This study shows that induction of miRNAs by P. gingivalis can modulate important host signaling responses.