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分析经典霍奇金淋巴瘤 L428 细胞系中 CCL5 的表达。

Analysis of CCL5 expression in classical Hodgkin's lymphoma L428 cell line.

机构信息

Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.

出版信息

Mol Med Rep. 2011 Sep-Oct;4(5):837-41. doi: 10.3892/mmr.2011.515. Epub 2011 Jun 22.

Abstract

CCL5 is one of the chemoattractant cytokines involved in inflammatory observed in both diffuse large B-cell lymphoma (DLBCL) and classical Hodgkin's lymphoma (CHL). However, the pathological effects of CCL5 remain unclear. To gain a better understanding of the role of CCL5 in CHL and DLBCL, we examined the expression of CCL5 in the CHL cell line L428 and the DLBCL cell lines Ly1 and Ly8, as well as its chemotactic effect on CD4+ T cells. CCL5 mRNA expression was detected by real-time quantitative RT-PCR. Intracellular CCL5 protein expression was analyzed using confocal microscopy, and CCL5 protein secretion was detected by ELISA. The chemotactic function of CCL5 was assessed using a Transwell coculture system, and the number of migrated CD4+ T cells was counted. Moreover, the p-iкBα and p65 levels of NF-кB signaling molecules in these lymphoma cell lines were detected by Western blotting. The results showed that CCL5 mRNA and protein expression in the L428 cells was significantly higher than in Ly1 and Ly8 cells (p<0.05). L428 cells secreted more CCL5 than the Ly1 or Ly8 cells, and the secreted CCL5 was capable of inducing CD4+ T cell migration. The expression levels of the NF-кB transcription factors p65 and p-iкBα were examined in these lymphoma cells. L428, Ly1 and Ly8 cells expressed similar levels of p65, while p-iкBα expression was higher in the L428 cells than in the Ly1 or Ly8 cells, indicating that a high CCL5 expression may be related to the increased activity of the NF-кB signaling pathway in L428 cells.

摘要

CCL5 是一种趋化因子细胞因子,参与弥漫性大 B 细胞淋巴瘤 (DLBCL) 和经典霍奇金淋巴瘤 (CHL) 中观察到的炎症反应。然而,CCL5 的病理作用尚不清楚。为了更好地了解 CCL5 在 CHL 和 DLBCL 中的作用,我们检测了 CHL 细胞系 L428 和 DLBCL 细胞系 Ly1 和 Ly8 中 CCL5 的表达及其对 CD4+T 细胞的趋化作用。通过实时定量 RT-PCR 检测 CCL5 mRNA 表达。使用共聚焦显微镜分析细胞内 CCL5 蛋白表达,通过 ELISA 检测 CCL5 蛋白分泌。使用 Transwell 共培养系统评估 CCL5 的趋化功能,并计数迁移的 CD4+T 细胞数量。此外,通过 Western blot 检测这些淋巴瘤细胞系中 NF-κB 信号分子的 p-iкBα 和 p65 水平。结果显示,L428 细胞中 CCL5 mRNA 和蛋白表达明显高于 Ly1 和 Ly8 细胞(p<0.05)。L428 细胞分泌的 CCL5 多于 Ly1 或 Ly8 细胞,分泌的 CCL5 能够诱导 CD4+T 细胞迁移。检测了这些淋巴瘤细胞中 NF-κB 转录因子 p65 和 p-iкBα 的表达水平。L428、Ly1 和 Ly8 细胞表达相似水平的 p65,而 L428 细胞中 p-iкBα 的表达高于 Ly1 或 Ly8 细胞,表明 CCL5 表达水平高可能与 L428 细胞中 NF-κB 信号通路活性增加有关。

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