红花黄色素通过 FAK 通路调控免疫耐受抑制肝癌进展。
Safflower Yellow Inhibits Progression of Hepatocellular Carcinoma by Modulating Immunological Tolerance via FAK Pathway.
机构信息
Department of General Surgery, First People's Hospital of Hangzhou Lin'an District, Hangzhou, 311300, China.
出版信息
Chin J Integr Med. 2024 Apr;30(4):339-347. doi: 10.1007/s11655-023-3705-1. Epub 2023 Nov 9.
OBJECTIVE
To explore the anti-tumor effect of safflower yellow (SY) against hepatocellular carcinoma (HCC) and the underlying potential mechanism.
METHODS
An in vitro model was established by mixing Luc-Hepa1-6 cells and CD3CD8 T cells, followed by adding programmed cell death protein 1 (PD-1) antibody (Anti-mPD-1) with or without SY. The apoptosis was detected by flow cytometry and the level of inflammatory cytokines was determined by enzyme-linked immunosorbent assay. The protein levels of programmed cell death 1 ligand 1 (PD-L1), chemokine ligand (CCL5), C-X-C motif chemokine ligand 10 (CXCL10) were measured by Western blot. An in situ animal model was established in mice followed by treatment with anti-mPD-1 with or without SY. Bioluminescence imaging was monitored with an AniView 100 imaging system. To establish the FAK-overexpressed Luc-Hepa1-6 cells, cells were transfected with adenovirus containing pcDNA3.1-FAK for 48 h.
RESULTS
The fluorescence intensity, apoptotic rate, release of inflammatory cytokines, and CCL5/CXCL10 secretion were dramatically facilitated by anti-mPD-1 (P<0.01), accompanied by an inactivation of PD-1/PD-L1 axis, which were extremely further enhanced by SY (P<0.05 or P<0.01). Increased fluorescence intensity, elevated percentage of CD3CD8 T cells, facilitated release of inflammatory cytokines, inactivated PD-1/PD-L1 axis, and increased CCL5/CXCL10 secretion were observed in Anti-mPD-1 treated mice (P<0.01), which were markedly enhanced by SY (P<0.05 or P<0.01). Furthermore, the enhanced effects of SY on inhibiting tumor cell growth, facilitating apoptosis and inflammatory cytokine releasing, suppressing the PD-1/PD-L1 axis, and inducing the CCL5/CXCL10 secretion in Anti-mPD-1 treated mixture of Luc-Hepa1-6 cells and CD3CD8 T cells were abolished by FAK overexpression (P<0.01).
CONCLUSION
SY inhibited the progression of HCC by mediating immunological tolerance through inhibiting FAK.
目的
探讨红花黄色素（SY）对肝癌（HCC）的抗肿瘤作用及其潜在机制。
方法
体外构建 Luc-Hepa1-6 细胞与 CD3CD8 T 细胞混合培养模型，加入程序性死亡蛋白 1（PD-1）抗体（Anti-mPD-1），并分别给予 SY 或不给予 SY，通过流式细胞术检测细胞凋亡情况，酶联免疫吸附法检测炎症因子水平，Western blot 法检测程序性死亡配体 1（PD-L1）、趋化因子配体（CCL5）、C-X-C 基元趋化因子配体 10（CXCL10）的蛋白水平。构建小鼠原位动物模型，分别给予 Anti-mPD-1 联合或不联合 SY 治疗，使用 AniView 100 成像系统进行生物发光成像监测。使用含有 pcDNA3.1-FAK 的腺病毒转染 Luc-Hepa1-6 细胞 48 h，构建 FAK 过表达 Luc-Hepa1-6 细胞。
结果
Anti-mPD-1 明显促进了荧光强度、细胞凋亡率、炎症因子释放和 CCL5/CXCL10 的分泌（P<0.01），同时抑制了 PD-1/PD-L1 轴，SY 进一步增强了上述作用（P<0.05 或 P<0.01）。Anti-mPD-1 治疗组小鼠的荧光强度增加、CD3CD8 T 细胞比例升高、炎症因子释放增加、PD-1/PD-L1 轴失活、CCL5/CXCL10 分泌增加（P<0.01），SY 明显增强了上述作用（P<0.05 或 P<0.01）。此外，FAK 过表达削弱了 SY 对 Anti-mPD-1 治疗的 Luc-Hepa1-6 细胞和 CD3CD8 T 细胞混合物的肿瘤细胞生长抑制、促进凋亡和炎症因子释放、抑制 PD-1/PD-L1 轴、诱导 CCL5/CXCL10 分泌的增强作用（P<0.01）。
结论
SY 通过抑制 FAK 介导的免疫耐受抑制 HCC 的进展。
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