Inhibition of Notch 1 receptor influenced the differentiation of Lin-CD45RA-dendritic cell precursors within ovarian carcinoma microenvironment.
Inhibition of Notch 1 receptor influenced the differentiation of Lin-CD45RA-dendritic cell precursors within ovarian carcinoma microenvironment.
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DOI：
10.1186/s12865-016-0150-3
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发表时间：
2016-06-04
期刊：
影响因子：
3
通讯作者：
Wan XY
中科院分区：
文献类型：
--
作者：
Qian XQ;Chen LL;Cheng Q;Tian Y;Luo XF;Wan XY
先前的证据表明，在卵巢癌微环境中，Lin-CD45RA-DC前体的分化先于质细胞树突状细胞（pDCs）而非髓细胞树突状细胞（mDCs）。然而，其机制尚不清楚。因此，我们研究Notch 1信号通路在Lin-CD45RA-DC前体分化中的作用。从足月分娩的脐带血中提取CD34+造血干细胞，分离Lin-CD45RA-DC前体并诱导成熟。通过Real-time PCR检测Lin-CD45RA-DC前体Notch1受体和配体的表达，并通过shRNA或γ-分泌酶抑制剂（GSI）下调Notch1受体和配体的表达。流式细胞术分析有或没有SKOV3培养上清的DCs亚群。ELISA法检测IL-12水平。在Lin-CD45RA-DC前体细胞中检测到Notch1受体和配体的表达。shRNA-Notch1慢病毒转染和GSI可下调Lin-CD45RA-DC前体的Notch1 mRNA表达。ShRNA介导的Notch 1敲除显著减少了质胞质树突状细胞（pDCs）的分化，但产生了更多的髓系树突状细胞（mDCs），这不会受到卵巢癌细胞系上清的影响。GSI对pDC的分化效果相同。不论有无SKOV3培养上清，Notch1敲除后IL-12的分泌均显著增加。Notch1是Lin-CD45RA-DC前体细胞向质细胞质树突状细胞（pDCs）分化的重要信号通路。这不会受到卵巢癌细胞系上清液的影响。
Previous evidence suggested that the differentiation of Lin-CD45RA-DC precursors were prior to plasmcytoid dendritic cells (pDCs) than myeloid dendritic cells (mDCs) within ovarian cancer microenvironment. However, the mechanism is still unclear. Therefore, we investigated the function of Notch 1 signal pathway in the differentiation of Lin-CD45RA-DC precursors. The CD34+ hematopoietic stem cells were extracted from umbilical cord blood in term parturition, and Lin-CD45RA-DC precusors were separated and induced mature. Expression of Notch1 receptor and ligands in Lin-CD45RA-DC precusors was detected by Real-time PCR and was down-regulated by shRNA or γ-secretase inhibitor (GSI). Flow cytometry was used to analyze the subset of DCs with or without SKOV3 culture supernatants. IL-12 level was detected by ELISA. Expression of Notch1 receptors and ligands were detected in Lin-CD45RA-DC precursor cells. The Notch1 mRNA in Lin-CD45RA-DC precursors can be down-regulated by shRNA-Notch1 lentivirus transfection and GSI. ShRNA mediated Notch 1 knock-down significantly differentiated less plasmcytoid dendritic cells (pDCs), but generated more myeloid dendritic cells (mDCs), and this would not be influenced by the supernatant of the ovarian carcinoma cell line. GSI had the same effect in the differentiation of pDC. The secretion of IL-12 significantly increased after Notch1 knock-down with or without SKOV3 culture supernatants. Notch1 is an important signaling pathway in the differentiation of Lin-CD45RA-DC precursor cells to plasmcytoid dendritic cells (pDCs). And this would not be affected by the supernatant of the ovarian carcinoma cell line.