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在 K562 细胞培养的条件培养基(CM)中培养的外周血单个核细胞(PBMCs)中,TGF-β和 IFN-γ的表达增加。

Increased Expression of TGF-β and IFN-γ in Peripheral Blood Mononuclear Cells (PBMCs) Cultured in Conditioned Medium (CM) of K562 Cell Culture.

机构信息

School of Life Sciences, B.S. Abdur Rahman Crescent Institute of Science and Technology, Vandalur, Chennai, Tamil Nadu, India 600048.

Translational Research Platform for Veterinary Biologicals, Central University Laboratory Building, TANUVAS, Madhavaram Milk Colony, Chennai, Tamil Nadu, India 600051.

出版信息

J Environ Pathol Toxicol Oncol. 2019;38(2):173-183. doi: 10.1615/JEnvironPatholToxicolOncol.2019029460.

Abstract

In the present study, we investigated the effects of conditioned media (CM) collected from the cancer cell lines (K562, MCF-7, and HeLa) on peripheral blood mononuclear cells (PBMCs) isolated from the healthy human blood. The soluble factors in the CM are probably responsible for the differential mRNA expressions of Foxp3, Helios, Neuropilin- 1 (NRP-1), and glycoprotein A repetitions predominant (GARP), along with IFN-γ and TGF-β in PBMCs cultured with cancer cells CM. The PBMCs cultured with CM of K562 showed increased expression of Foxp3, Helios, NRP-1, GARP, IFN-γ, and TGF-β compared to PBMCs cultured with CM of MCF-7 and HeLa cells. In addition, the intracellular staining on PBMCs cultured with CM from cell lines were also evaluated for CD4, CD25, Foxp3, Helios, and NRP-1 by multicolor flow cytometry. The expression of CD4+CD25+Foxp3+, CD4+Helios+Foxp3+ and CD+NRP-1+Foxp3+ showed retarded cell population compared to control PBMCs. Our data suggest that soluble factors in CM of cancer cells may trigger the immune response in PBMCs resulting in a systematic response. Further research could lead to the identification of specific soluble factors that are involved in trafficking of cells into the immune cascades, which could be a safe and promising strategy for targeting human cancers.

摘要

在本研究中,我们研究了来自癌细胞系(K562、MCF-7 和 HeLa)的条件培养基(CM)对来自健康人血液的外周血单个核细胞(PBMC)的影响。CM 中的可溶性因子可能导致 Foxp3、Helios、神经纤毛蛋白-1(NRP-1)和糖蛋白 A 重复为主(GARP)的差异 mRNA 表达以及 IFN-γ和 TGF-β在与癌细胞 CM 共培养的 PBMC 中。与与 MCF-7 和 HeLa 细胞 CM 共培养的 PBMC 相比,与 K562 细胞 CM 共培养的 PBMC 表现出 Foxp3、Helios、NRP-1、GARP、IFN-γ和 TGF-β的表达增加。此外,还通过多色流式细胞术评估了 PBMC 与来自细胞系的 CM 共培养后的细胞内染色,用于 CD4、CD25、Foxp3、Helios 和 NRP-1。与对照 PBMC 相比,CD4+CD25+Foxp3+、CD4+Helios+Foxp3+和 CD+NRP-1+Foxp3+的表达显示出延迟的细胞群体。我们的数据表明,癌细胞 CM 中的可溶性因子可能引发 PBMC 中的免疫反应,导致系统反应。进一步的研究可能导致鉴定出参与细胞进入免疫级联的特定可溶性因子,这可能是针对人类癌症的安全且有前途的策略。

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