胶束莪术醇对卵巢癌相关巨噬细胞极化的影响
Effect of micellar curcumol on polarization of macrophages associated with ovarian cancer
唐勤 1王晶 2陈冰 3汪生 4张敏敏 2张梦媛 2吴强5
作者信息
- 1. 安徽医科大学基础医学院病理学教研室,合肥 230032
- 2. 安徽医科大学第一附属医院妇产科,合肥 230022
- 3. 合肥工业大学化学与材料科学学院,合肥 230009
- 4. 安徽医科大学科研实验中心,合肥 230032
- 5. 安徽医科大学基础医学院病理学教研室,合肥 230032;安徽医科大学第一附属医院病理科,合肥 230022
- 折叠
摘要
目的 探讨纳米材料胶束莪术醇(MC)通过促进卵巢癌腹水M2型巨噬细胞向M1型极化调控卵巢癌免疫微环境的机制.方法 ① 小鼠分组后,用鼠卵巢癌细胞株ID8构建卵巢癌腹水模型,观察体质量变化,收集肿瘤组织和腹水.流式细胞术检测肿瘤组织和腹水巨噬细胞CD86和CD206表达;Western blot法检测蛋白激酶B(PKB/Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)表达.② 诱导人单核细胞白血病(THP-1)细胞转化M2型巨噬细胞(THP-1 M2φ),用10μg/ml的MC处理,流式法检测凋亡率;qRT-PCR法检测巨噬细胞甘露糖受体(CD206)、转化生长因子-β(TGF-β)和白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)的mRNA表达;流式法检测CD86和CD206表达;Western blot法检测Akt/mTOR表达.结果 ① 体内实验:给药后,MC组鼠平均体质量低于对照组;MC组肿瘤组织和腹水巨噬细胞CD206表达降低,CD86表达上调;MC组腹水Akt、mTOR磷酸化水平降低.② 体外实验:用药前后THP-1 M2φ 凋亡无差异;MC组CD206、TGF-β的mRNA表达及CD206蛋白表达明显低于对照组,同时IL-1β、TNF-α的mRNA和CD86蛋白表达明显高于对照组;MC组Akt、mTOR的磷酸化水平降低.结论 MC促进腹水巨噬细胞向M1极化调控卵巢癌免疫微环境,其机制可能与Akt/mTOR通路有关.
Abstract
Objective To investigate the mechanism of micellar curcumol (MC) regulating the immune microenvi-ronment of ovarian cancer by promoting the polarization of M2-type macrophages to M1-type in ovarian cancer asci-tes.Methods ① After the mice were divided into groups, a mouse ovarian cancer ascites model was constructed by using the mouse ovarian cancer cell line ID8.Then weight changes were observed, tumor tissue and ascites were collected.The expression of CD86 and CD206 on macrophages of the tumor tissue and ascites was detected by flow cytometry.The expression of protein kinase B (PKB/Akt)/mammalian target of rapamycin (mTOR) was detected by Western blot.②A human monocytic leukemia cell line (THP-1) was induced to transform into M2 macrophage (THP-1 M2 macrophage) in vitro, and then treated with 10μg/ml MC.The apoptosis was detected by flow cytom-etry.The mRNA levels of macrophage mannose receptor (CD206), transforming growth factor-β(TGF-β), inter-leukin (IL)-1β and tumor necrosis factor-α (TNF-α) were detected by qRT-PCR.The expression of CD86 and CD206 was detected by flow cytometry, and Akt/mTOR expression and phosphorylation was detected by Western blot.Results ① In vitro study showed that the average body weight of the MC group was lower than that of the control group.Compared with the control group, CD206 expression of macrophages decreased in tumor tissue and ascites in the MC group, while the expression of CD86 increased.The Akt and mTOR phosphorylation level of mac-rophages in the MC group's ascites was lower than that in control group.②In vivo study showed that there was no difference in apoptosis rate among the groups detected by flow cytometry.The mRNA expression level of CD206, TGF-β and the protein expression level of CD206 in MC group were significantly lower than those in the control group, while the mRNA expression of IL-1β, TNF-α and the protein expression level of CD86 were significantly higher than those in the control group.Compared with the control group, the phosphorylation level of Akt and mTOR in the MC group decreased.Conclusion MC promotes M1 polarization of macrophages in ascites to regulate the immune microenvironment of ovarian cancer, which may be related to the Akt/mTOR pathway.
关键词
卵巢癌/免疫微环境/巨噬细胞/MC/Akt/mTORKey words
ovarian cancer/immune microenvironment/macrophages/MC/Akt/mTOR引用本文复制引用
基金项目
国家自然科学基金(82104633)
安徽高校研究生科学研究项目(YJS20210282)
安徽高校自然科学研究项目(2023AH053293)
安徽高校自然科学研究项目(KJ2020A0200)
出版年
2024
国家科技期刊平台
NSTL
万方数据