[1]王佳勇 朱晓俊 王素娴 高薇 严自强**.黄精多糖调控巨噬细胞极化抑制食管癌细胞Eca109迁移侵袭的作用机制[J].现代中医药,2024,(06):088-95.[doi:10.13424/j.cnki.mtcm.2024.06.016]
　WANG Jiayong ZHU Xiaojun WANG Suxian GAO Wei YAN Ziqiang.Mechanism of Polygonatum Sibiricum Polysaccharides RegulatingMacrophage Polarization and Inhibiting Migration andInvasion of Esophageal Cancer Cell Line Eca109[J].Modern Traditional Chinese Medicine,2024,(06):088-95.[doi:10.13424/j.cnki.mtcm.2024.06.016]
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黄精多糖调控巨噬细胞极化抑制食管癌细胞Eca109迁移侵袭的作用机制()
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《现代中医药》[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2024年06期
页码:
088-95
栏目:
出版日期:
2024-11-20

文章信息/Info

Title:
Mechanism of Polygonatum Sibiricum Polysaccharides RegulatingMacrophage Polarization and Inhibiting Migration andInvasion of Esophageal Cancer Cell Line Eca109
文章编号:
1672-0571(2024)04-0079-04
作者:
王佳勇 朱晓俊 王素娴 高薇 严自强**
海军军医大学附属长海医院，上海 200433
Author(s):
WANG Jiayong ZHU Xiaojun WANG Suxian GAO Wei YAN Ziqiang
Naval Medical University Affiliated Changhai Hospital, Shanghai 200433,China
关键词:
关键词：黄精多糖; 食管癌; 肿瘤相关巨噬细胞极化; 侵袭; 迁移
Keywords:
Key words:Polygonatum sibiricum polysaccharide;  Esophageal cancer;  Tumor associated macrophage polarization;  Attack;  Transfer
分类号:
R571
DOI:
10.13424/j.cnki.mtcm.2024.06.016
文献标志码:
A
摘要:
摘 要：目的 探究黄精多糖(Polygonatum sibiricum polysaccharides，PSP)调控巨噬细胞极化对食管癌细胞Eca109侵袭迁移的影响及作用机制。方法 采用细胞计数试剂盒（Cell Counting Kit-8，CCK-8）法检测PSP对巨噬细胞RAW264.7存活率的影响；白细胞介素4（Interleukin 4，IL-4）（20 ng·mL-1）/白细胞介素13（Interleukin 13，IL-13）（20 ng·mL-1）诱导RAW264.7细胞极化为M2型巨噬细胞，并给予PSP低、中、高（50、100、200 μg·mL-1）干预24 h，并收集细胞培养上清制备条件培养基。采用反转录-聚合酶链反应（reverse transcription-polymerase chain reaction，RT-PCR）检测M2型巨噬细胞标志物精氨酸酶-1（Arginase-1，Arg-1）、甘露糖受体1（mannose receptor 1，CD206）、类几丁质酶3样分子3（Chitinase 3-like 3，Ym-1）mRNA的水平；采用流式细胞术检测M2型巨噬细胞表面抗原CD206的表达；划痕实验检测条件培养基对Eca109细胞迁移能力的影响；Transwell实验检测条件培养基对Eca109细胞侵袭能力的影响；采用Western blot 实验检测细胞内蛋白酪氨酸激酶3（Janus kinase 3，JAK3）、磷酸化蛋白酪氨酸激酶3（Phospho-Janus kinase 3，p-JAK3）、信号转导及转录激活蛋白6（Signal transducer and activator of transcription 6，STAT6）、磷酸化信号转导及转录激活蛋白6（Phospho-signal transducer and activator of transcription 6，p-STAT6）蛋白的表达。结果 PSP在浓度低于200 μg·mL-1时，对RAW264.7细胞存活率没有显著影响（P≥005）。PSP（50、100、200 μg·mL-1）呈剂量依赖降低CD206+细胞比例，减少Arg-1、CD206、Ym-1 mRNA的水平（P＜0.05）。PSP巨噬细胞条件培养基显著抑制Eca109细胞的侵袭迁移能力。PSP（50、100、200 μg·mL-1）明显降低p-JAK3/JAK3和p-STAT6/STAT6的蛋白表达比值（P<0.05）。结论 PSP可以调控巨噬细胞M2极化抑制食管癌细胞Eca109侵袭迁移，其作用机制可能与抑制JAK3/STAT6信号通路有关。
Abstract:
Abstract：Objective To investigate the effect and mechanism of Polygonatum sibiricum polysaccharides (PSP) on macrophage polarization and invasion and migration of esophageal cancer cell line Eca109. Methods The Cell Counting Kit-8 (CCK-8) assay was used to detect the effect of PSP on the survival rate of RAW264.7 macrophages; Interleukin-4 (IL-4) (20 ng·mL-1)/interleukin-13 (IL-13) (20 ng·mL-1) induced polarization of RAW264.7 cells into M2 macrophages, and PSP low, medium, and high (50, 100, 200 μg·mL-1) interventions were administered for 24 hours. Cell culture supernatant was collected to prepare conditioned medium. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA levels of M2 macrophage markers Arginase-1 (Arg-1), mannose receptor 1 (CD206), and Chitinase 3-like 3 (Ym-1); Use flow cytometry to detect the expression of M2 macrophage surface antigen CD206; The effect of scratch test conditions and culture medium on the migration ability of Eca109 cells; Transwell experiment detects the effect of conditioned medium on the invasive ability of Eca109 cells; Western blot assay was used to detect the expression of protein tyrosine kinase 3 (JAK3), phosphorylated protein tyrosine kinase 3 (p-JAK3), signal transducer and activator of transcription 6 (STAT6), and phosphorylated signal transducer and activator of transcription 6 (p-STAT6) proteins in cells. Results The results showed that PSP had no significant effect on the survival rate of RAW264.7 cells at concentrations below 200 μg·mL-1 (P≥0.05).PSP (50, 100, 200 μg·mL-1) showed a dose-dependent decrease in the proportion of CD206+ cells and reduced the levels of Arg-1, CD206, and Ym-1 mRNA (P<0.05). PSP macrophage conditioned medium significantly inhibits the invasion and migration ability of Eca109 cells. PSP (50, 100, 200 μg·mL-1) significantly reduced the protein expression ratios of p-JAK3/JAK3 and p-STAT6/STAT6 (P<0.05). Conclusion PSP can regulate macrophage M2 polarization and inhibit the invasion and migration of esophageal cancer cell line Eca109, and its mechanism of action may be related to the inhibition of the JAK3/STAT6 signaling pathway.

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备注/Memo

备注/Memo:
基金项目：国家自然科学基金项目（81703882）；上海市药学会项目（2019-YY-09）
更新日期/Last Update: 2024-11-21