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缺氧条件下调节性T细胞和吲哚胺2,3-双加氧酶对人上皮性卵巢癌细胞的影响。

Effects of Treg cells and IDO on human epithelial ovarian cancer cells under hypoxic conditions.

作者信息

Liu Jun, Zhang Haiyan, Jia Luoqi, Sun Hong

机构信息

Department of Gynecology, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200090, P.R. China.

出版信息

Mol Med Rep. 2015 Mar;11(3):1708-14. doi: 10.3892/mmr.2014.2893. Epub 2014 Nov 7.

Abstract

The aim of the present study was to explore the effect of hypoxia on ovarian cancer. A total of 6 samples were analyzed: SKOV3‑IP cells (ovarian cancer cell line); SKOV3‑IP and regulatory T (Treg) cells; SKOV3‑IP and cytotoxic T lymphocytes (CTLs); SKOV3‑IP and natural killer (NK) cells; SKOV3‑IP co-cultured with CTLs and Treg cells; and SKOV3‑IP co-cultured with Treg cells and NK cells. The expression of indoleamine 2,3‑dioxygenase (IDO) was detected by reverse transcription-polymerase chain reaction (RT‑PCR) and western blot analysis. An enzyme‑linked immunosorbent assay (ELISA) was used to detect the concentration of transforming growth factor‑β (TGF‑β), interferon‑γ (IFN‑γ), interleukin‑2 (IL‑2), interleukin‑10 (IL‑10), and perforin. Moreover, ovarian cancer cell apoptosis and invasive ability were examined using flow cytometry and a Transwell chamber assay. IDO expression was significantly reduced in hypoxia and enhanced by Treg cells. Treg cells inhibited the IL‑2, IFN‑γ and perforin secretion, and significantly (P<0.05) increased the IL‑10 and TGF‑β levels. The effects of Treg cells were enhanced with prolongation of the cell exposure to hypoxic conditions. In addition, Treg cells attenuated the promotive effect of CTLs and NK cells on cancer cell apoptosis. In addition, Treg cells significantly increased the SKOV3‑IP invasive ability (P=0.00109) under hypoxic conditions. Our results suggest that IDO and Treg cells may serve as important therapeutic targets for patients with ovarian cancer.

摘要

本研究的目的是探讨缺氧对卵巢癌的影响。共分析了6个样本:SKOV3-IP细胞(卵巢癌细胞系);SKOV3-IP细胞与调节性T(Treg)细胞;SKOV3-IP细胞与细胞毒性T淋巴细胞(CTLs);SKOV3-IP细胞与自然杀伤(NK)细胞;SKOV3-IP细胞与CTLs和Treg细胞共培养;以及SKOV3-IP细胞与Treg细胞和NK细胞共培养。通过逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹分析检测吲哚胺2,3-双加氧酶(IDO)的表达。采用酶联免疫吸附测定(ELISA)检测转化生长因子-β(TGF-β)、干扰素-γ(IFN-γ)、白细胞介素-2(IL-2)、白细胞介素-10(IL-10)和穿孔素的浓度。此外,使用流式细胞术和Transwell小室测定法检测卵巢癌细胞凋亡和侵袭能力。缺氧时IDO表达显著降低,Treg细胞可增强其表达。Treg细胞抑制IL-2、IFN-γ和穿孔素的分泌,并显著(P<0.05)提高IL-10和TGF-β水平。随着细胞暴露于缺氧条件时间的延长,Treg细胞的作用增强。此外,Treg细胞减弱了CTLs和NK细胞对癌细胞凋亡的促进作用。此外,在缺氧条件下,Treg细胞显著提高了SKOV3-IP细胞的侵袭能力(P=0.00109)。我们的结果表明,IDO和Treg细胞可能是卵巢癌患者重要的治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad91/4270340/413107745c62/MMR-11-03-1708-g00.jpg

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