[1]李浩,戎帅,刘连涛,等.鸦胆子苦醇通过CC趋化因子配体2-C-C趋化因子受体2信号通路对骨肉瘤细胞增殖、凋亡和免疫逃逸的影响[J].中国中医骨伤科杂志,2023,31(12):1-6+14.[doi:10.20085/j.cnki.issn1005-0205.231201]
 LI Hao,RONG Shuai,LIU Liantao,et al.Effect of Brusatol on the Proliferation,Apoptosis and Immune Escape of Osteosarcoma Cells by CCL2-CCR2 Signal Pathway[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2023,31(12):1-6+14.[doi:10.20085/j.cnki.issn1005-0205.231201]
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鸦胆子苦醇通过CC趋化因子配体2-C-C趋化因子受体2信号通路对骨肉瘤细胞增殖、凋亡和免疫逃逸的影响()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第31卷
期数:
2023年12期
页码:
1-6+14
栏目:
实验研究
出版日期:
2023-12-15

文章信息/Info

Title:
Effect of Brusatol on the Proliferation,Apoptosis and Immune Escape of Osteosarcoma Cells by CCL2-CCR2 Signal Pathway
文章编号:
1005-0205(2023)12-0001-06
作者:
李浩1戎帅1刘连涛1李克伟1△
1石家庄市第三医院小儿骨科(石家庄,050011)
Author(s):
LI Hao1RONG Shuai1LIU Liantao1LI Kewei1△
1Department of Pediatric Orthopedics,Shijiazhuang Third Hospital,Shijiazhuang 050011,China.
关键词:
鸦胆子苦醇 骨肉瘤 增殖 凋亡 免疫逃逸 CC趋化因子配体2-C-C趋化因子受体2信号通路
Keywords:
brusatol osteosarcoma proliferation apoptosis immune escape CCL2-CCR2 signal pathway
分类号:
R-33
DOI:
10.20085/j.cnki.issn1005-0205.231201
文献标志码:
A
摘要:
目的:探讨鸦胆子苦醇(BRU)调节CC趋化因子配体2(CCL2)-C-C趋化因子受体2(CCR2)信号通路对骨肉瘤(OS)细胞增殖、凋亡和免疫逃逸的影响。方法:将骨肉瘤细胞系MG-63分为MG-63组(未处理的MG-63细胞)、L-BRU组(25 nmol/L鸦胆子苦醇处理MG-63细胞)、M-BRU组(50 nmol/L鸦胆子苦醇处理MG-63细胞)、H-BRU组(75 nmol/L鸦胆子苦醇处理MG-63细胞)、GW0742组(1 μmol/L CCL2-CCR2信号通路激活剂GW0742处理MG-63细胞)、H-BRU+GW0742组(1 μmol/L GW0742和 75 nmol/L鸦胆子苦醇处理MG-63细胞),MTT法检测鸦胆子苦醇对正常人成骨细胞系hFOB1.19细胞的毒性,CCK8法检测MG-63细胞增殖,流式细胞仪检测MG-63细胞、CD8+T细胞凋亡,ELISA法检测外周血单个核细胞(PBMC)上清液IFN-γ、TNF-α水平,Western Blot法检测CCL2-CCR2通路蛋白以及凋亡相关蛋白水平。结果:0~500 nmol/L鸦胆子苦醇对hFOB1.19细胞无明显毒性影响; 与MG-63组相比,L-BRU组、M-BRU组、H-BRU组光密度(OD)值,Bcl-2、CCL2、CCR2、PD-L1蛋白水平及CD8+T细胞的凋亡率均显著降低(P<0.05),凋亡率,IFN-γ、TNF-α水平及Bax、Cleaved-Caspase-3蛋白水平均显著升高(P<0.05),而GW0742组以上指标趋势相反; 与H-BRU组相比,H-BRU+GW0742组光密度值,Bcl-2、CCL2、CCR2、PD-L1蛋白水平及CD8+T细胞的凋亡率均显著增加(P<0.05),凋亡率,IFN-γ、TNF-α水平及Bax、Cleaved-Caspase-3蛋白水平均显著降低(P<0.05)。结论:鸦胆子苦醇可能通过抑制CCL2-CCR2信号通路对骨肉瘤细胞增殖、凋亡和免疫逃逸产生影响。
Abstract:
Objective:To investigate the effect of brusatol(BRU)on the proliferation,apoptosis and immune escape of osteosarcoma(OS)cells by regulating CC chemokine ligand 2(CCL2)-C-C chemokine receptor 2(CCR2)signal pathway.Methods:The osteosarcoma cell line MG-63 were divided into MG-63 group(untreated MG-63 cells),L-BRU group(MG-63 cells treated with 25 nmol/L BRU),M-BRU group(MG-63 cells treated with 50 nmol/L BRU),H-BRU group(MG-63 cells treated with 75 nmol/L BRU),GW0742 group(MG-63 cells treated with 1 μmol/L CCL2-CCR2 signal pathway activator GW0742),and H-BRU+GW0742 group(MG-63 cells treated with 1 μmol/L GW0742 and 75 nmol/L BRU),the MTT method was used to detect the toxicity of BRU to normal human osteoblast line hFOB1.19 cells,CCK8 method was applied to detect MG-63 cell proliferation,flow cytometry was applied to detect apoptosis of MG-63 cells and CD8+T cells,ELISA method was applied to detect the levels of IFN-γ and TNF-α in PBMC supernatant,and Western Blot was applied to detect the levels of CCL2-CCR2 pathway proteins and apoptosis-related proteins.Results:0500 nmol/L BRU had no obvious toxic effect on hFOB1.19 cells.Compared with MG-63 group,the OD value,Bcl-2,CCL2,CCR2,PD-L1 protein levels and the apoptosis rate of CD8+T cells in L-BRU group,M-BRU group and H-BRU group were obviously lower(P<0.05),and the apoptosis rate,IFN-γ,TNF-α levels,Bax,Cleaved-Caspase-3 protein levels were obviously higher(P<0.05),the trend of the above indicators in GW0742 group was opposite.Compared with H-BRU group,the OD value,Bcl-2,CCL2,CCR2,PD-L1 protein levels and the apoptosis rate of CD8+T cells in H-BRU+GW0742 group were obviously higher(P<0.05),the apoptosis rate,IFN-γ,TNF-α levels,Bax,Cleaved-Caspase-3 protein levels were obviously lower(P<0.05).Conclusion:BRU may affect the proliferation,apoptosis and immune escape of osteosarcoma cells by inhibiting CCL2-CCR2 signal pathway.

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备注/Memo

备注/Memo:
基金项目:河北省医学科学研究重点课题计划项目(20191425)
通信作者 E-mail:10180737@qq.com
更新日期/Last Update: 2023-12-10