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二甲基亚砜诱导HL-60细胞分化过程中YKL-40(CHI3L1)的细胞内积累与分泌

Intracellular Accumulation and Secretion of YKL-40 (CHI3L1) in the Course of DMSO-Induced HL-60 Cell Differentiation.

作者信息

Jatczak-Pawlik Izabela, Ewiak-Paszyńska Alicja, Domowicz Małgorzata, Jurewicz Anna, Stasiołek Mariusz

机构信息

Department of Neurology, Medical University of Lodz, Kosciuszki Street 4, 90-419 Lodz, Poland.

出版信息

Pharmaceuticals (Basel). 2024 Mar 29;17(4):443. doi: 10.3390/ph17040443.

Abstract

YKL-40 (CHI3L1) is a matrix glycoprotein stored in human neutrophil-specific granules and released upon activation. While it is implicated in inflammation, cancer progression, and cell differentiation, its exact physiological role remains unclear. This study investigated the intracellular expression and secretion of YKL-40 by untreated and DMSO-treated HL-60 cells in association with surface expression of CD11b and CD66b throughout the differentiation process (up to 120 h). Secreted YKL-40 protein and mRNA levels of YKL-40, CD66b, and CD11b were measured by ELISA and quantitative RT-PCR, respectively. The intracellular YKL-40 and surface CD11b and CD66b expression were assessed by flow cytometry. A significant increase in CD11b expression confirmed DMSO-induced differentiation of HL-60 cells. Upon DMSO stimulation, YKL-40 mRNA expression increased in a time-dependent manner, unlike CD66b. The lack of CD66b (a granulocyte maturation and activation marker) on the surface of HL-60 cells might suggest that DMSO treatment did not induce full maturation or activation. The intracellular YKL-40 protein expression was increasing up to 96 h of DMSO treatment and then declined. YKL-40 secretion into the culture medium was detectable only at later time points (96 and 120 h), which was correlated with a decreased proliferation of DMSO-treated HL-60 cells. These findings suggest sequential changes in YKL-40 production and secretion during DMSO-induced differentiation of HL-60 cells and might contribute to a better understanding of YKL-40's involvement in both physiological processes and disease development, including multiple sclerosis.

摘要

YKL-40(CHI3L1)是一种储存于人类嗜中性粒细胞特异性颗粒中并在激活后释放的基质糖蛋白。虽然它与炎症、癌症进展和细胞分化有关,但其确切的生理作用仍不清楚。本研究调查了未处理和经二甲亚砜(DMSO)处理的HL-60细胞在整个分化过程(长达120小时)中YKL-40的细胞内表达和分泌情况,以及CD11b和CD66b的表面表达情况。分别通过酶联免疫吸附测定(ELISA)和定量逆转录聚合酶链反应(RT-PCR)检测分泌的YKL-40蛋白以及YKL-40、CD66b和CD11b的mRNA水平。通过流式细胞术评估细胞内YKL-40以及表面CD11b和CD66b的表达。CD11b表达的显著增加证实了DMSO诱导的HL-60细胞分化。在DMSO刺激下,与CD66b不同,YKL-40 mRNA表达呈时间依赖性增加。HL-60细胞表面缺乏CD66b(一种粒细胞成熟和激活标志物)可能表明DMSO处理未诱导完全成熟或激活。细胞内YKL-40蛋白表达在DMSO处理96小时前持续增加,随后下降。仅在后期时间点(96和120小时)可检测到YKL-40分泌到培养基中,这与经DMSO处理的HL-60细胞增殖减少相关。这些发现表明在DMSO诱导HL-60细胞分化过程中YKL-40产生和分泌的顺序变化,可能有助于更好地理解YKL-40在生理过程和疾病发展(包括多发性硬化症)中的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7042/11053806/6c1f75420699/pharmaceuticals-17-00443-g001.jpg

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