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Effect of siRNA-mediated downregulation of VEGF in Tca8113 cells on the activity of monocyte-derived dendritic cells

机译:siRNA介导的Tca8113细胞VEGF下调对单核细胞来源树突状细胞活性的影响

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摘要

Vascular endothelial growth factor (VEGF) is a tumor angiogenesis factor that is important in immune regulation. In our previous study, we found that VEGF expression in the peripheral blood and neoplasm nest from patients with oral squamous cell carcinoma (OSCC) was positively correlated with the course of disease, while an inverse correlation between VEGF expression and dendritic cells (DCs) was identified in the peripheral blood. Therefore, in the present study, we investigated whether inhibition of human VEGF in the human tongue carcinoma cell line Tca8113 had effects on the activity of monocyte-derived DCs. We knocked down the expression of human VEGF in Tca8113 cells using the small interfering RNA (siRNA) technique. Tca8113 cells pre-transfected with siRNA targeting VEGF were co-cultured with monocyte‑derived immature and mature DCs. Cell proliferation was evaluated by a WST-8 assay. Cell apoptosis, cell cycle and cell phenotypes were determined by flow cytometry. The data revealed that downregulation of the human VEGF significantly inhibited the proliferation of Tca8113 cells and increased apoptosis. Inhibition of human VEGF arrested the cell cycle of Tca8113 cells at the G0/G1 phase. Our results showed that the co-culture of DCs with Tca8113 cells markedly inhibited the expression of the mature markers of DCs including HLA-DR, CD80, CD86, CD40 and CD1a, as well as the immature marker CD83, while inhibition of human VEGF in Tca8113 cells significantly reversed these effects. Therefore, human VEGF in Tca8113 cells may not only regulate the cell proliferation and apoptosis of oral squamous cell carcinoma cells, but may also inhibit DC maturation.
机译:血管内皮生长因子(VEGF)是一种肿瘤血管生成因子,在免疫调节中很重要。在我们以前的研究中,我们发现口腔鳞状细胞癌(OSCC)患者外周血和肿瘤巢中的VEGF表达与疾病进程呈正相关,而VEGF表达与树突状细胞(DC)之间呈负相关。在外周血中鉴定。因此,在本研究中,我们研究了在人舌癌细胞系Tca8113中抑制人VEGF是否对单核细胞衍生DC的活性有影响。我们使用小干扰RNA(siRNA)技术敲低了Tca8113细胞中人VEGF的表达。用针对VEGF的siRNA预转染的Tca8113细胞与单核细胞来源的未成熟DC和成熟DC共培养。通过WST-8测定评估细胞增殖。通过流式细胞术确定细胞凋亡,细胞周期和细胞表型。数据显示人VEGF的下调显着抑制了Tca8113细胞的增殖并增加了细胞凋亡。人VEGF的抑制使Tca8113细胞的细胞周期停滞在G0 / G1期。我们的结果表明,DC与Tca8113细胞共培养可显着抑制DC的成熟标记物HLA-DR,CD80,CD86,CD40和CD1a以及未成熟的标记CD83的表达,同时抑制人VEGF的表达。 Tca8113细胞显着逆转了这些作用。因此,Tca8113细胞中的人VEGF不仅可以调节口腔鳞状细胞癌细胞的增殖和凋亡,而且还可以抑制DC成熟。

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