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健脾解毒方通过活性氧诱导的不完全自噬抑制结肠癌细胞增殖,并降低 PD-L1 表达。

Jianpi Jiedu Recipe Inhibits Proliferation through Reactive Oxygen Species-Induced Incomplete Autophagy and Reduces PD-L1 Expression in Colon Cancer.

机构信息

Yancheng TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Yancheng, Jiangsu, China.

Yancheng TCM Hospital, Yancheng, Jiangsu, China.

出版信息

Integr Cancer Ther. 2024 Jan-Dec;23:15347354241268064. doi: 10.1177/15347354241268064.

DOI:10.1177/15347354241268064
PMID:39155544
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11331576/
Abstract

BACKGROUND

Jianpi Jiedu Recipe has been used to treat digestive tract tumors in China since ancient times, and its reliability has been proven by clinical research. Currently, the specific biological mechanism of JPJDR in treating tumors is unclear.

METHODOLOGY

CCK-8 assay was used to detect cell viability. Clone formation assay and EdU assay were used to detect cell proliferation potential. DCFH-DA probe and JC-1 probe were used to detect total intracellular reactive oxygen species and mitochondrial membrane potential, respectively. Western blotting and immunofluorescence were used to detect protein expression level and subcellular localization of cells. The RFP-GFP-LC3B reporter system was used to observe the type of autophagy in cells. The xenograft tumor model was used to study the therapeutic effect of JPJDR in vivo.

RESULTS

JPJDR has an excellent inhibitory effect on various colorectal cancer cells and effectively reduces the proliferation ability of HT29 cells. After treatment with JPJDR, the amount of reactive oxygen species in HT29 cells increased significantly, and the mitochondrial membrane potential decreased. JPJDR induced the accumulation of autophagosomes in HT29 cells and was shown to be incomplete autophagy. At the same time, JPJDR reduced the expression of PD-L1. Meanwhile, JPJDR can exert an excellent therapeutic effect in xenograft tumor mice.

CONCLUSION

JPJDR is a low-toxicity and effective anti-tumor agent that can effectively treat colon cancer in vitro and in vivo. Its mechanism may be inducing mitochondrial dysfunction and incomplete autophagy injury to inhibit the proliferation of colon cancer cells.

摘要

背景

健脾解毒方自古代起就被用于治疗消化道肿瘤,其疗效已被临床研究所证实。目前,健脾解毒方治疗肿瘤的具体生物学机制尚不清楚。

方法

使用 CCK-8 法检测细胞活力。克隆形成实验和 EdU 实验检测细胞增殖能力。DCFH-DA 探针和 JC-1 探针分别检测细胞内总活性氧和线粒体膜电位。Western blot 和免疫荧光检测细胞内蛋白表达水平和亚细胞定位。使用 RFP-GFP-LC3B 报告系统观察细胞自噬类型。利用异种移植肿瘤模型研究 JPJDR 在体内的治疗效果。

结果

健脾解毒方对多种结直肠癌细胞具有优异的抑制作用,能有效降低 HT29 细胞的增殖能力。经 JPJDR 处理后,HT29 细胞内活性氧含量明显增加,线粒体膜电位降低。JPJDR 诱导 HT29 细胞中自噬体的积累,表现为不完全自噬。同时,JPJDR 降低了 PD-L1 的表达。此外,JPJDR 可在异种移植肿瘤小鼠中发挥出色的治疗效果。

结论

健脾解毒方是一种低毒、高效的抗肿瘤药物,可有效治疗结直肠肿瘤的体内外模型。其作用机制可能是通过诱导线粒体功能障碍和不完全自噬损伤来抑制结肠癌细胞的增殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/63ad7fa180f0/10.1177_15347354241268064-fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/f7d5b3fcb10d/10.1177_15347354241268064-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/8b6bced8d5c6/10.1177_15347354241268064-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/f64140f71fbd/10.1177_15347354241268064-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/699d6f469e56/10.1177_15347354241268064-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/fa1b5a1f9b8b/10.1177_15347354241268064-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/5f3f4c0700b8/10.1177_15347354241268064-fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/63ad7fa180f0/10.1177_15347354241268064-fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/f7d5b3fcb10d/10.1177_15347354241268064-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/8b6bced8d5c6/10.1177_15347354241268064-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/f64140f71fbd/10.1177_15347354241268064-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/699d6f469e56/10.1177_15347354241268064-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/fa1b5a1f9b8b/10.1177_15347354241268064-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/5f3f4c0700b8/10.1177_15347354241268064-fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3427/11331576/63ad7fa180f0/10.1177_15347354241268064-fig7.jpg

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