Huoluo Xiaolingdan Suppresses Triple-negative Breast Cancer in Mice by Regulating TCF1+ CD8+ Stem Cell-like T Cells Infiltration
10.13422/j.cnki.syfjx.20242021
- VernacularTitle:活络效灵丹调控TCF1+CD8+干细胞样T细胞肿瘤浸润抑制小鼠三阴乳腺癌
- Author:
Bo LUO
1
;
Qu ZHANG
1
;
Yujie SUN
2
;
Lin LIU
2
;
Lan ZENG
2
Author Information
1. Radiotherapy Center， Hubei Cancer Hospital， Wuhan 430079， China
2. School of Chinese Medicine， Hubei University of Chinese Medicine， Wuhan 430065， China
- Publication Type:Journal Article
- Keywords:
Huoluo Xiaolingdan;
breast cancer;
triple-negative breast cancer;
CD8+ T lymphocytes;
anti-tumor immunity;
stem cell
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(4):108-115
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the inhibitory effect of Huoluo Xiaolingdan on triple negative breast cancer （TNBC） in mice through its regulation of TCF1+CD8+ stem cell-like T cells infiltration. MethodsA mouse model of TNBC was established and the mice were randomly divided into the model group， low-dose （3.9 g·kg-1）， medium-dose （7.8 g·kg-1） and high-dose （15.6 g·kg-1） Huoluo Xiaolingdan groups， and anti-PD-1 antibody treatment group. Each group was given a dose of 0.01 mL·g-1， while the model group and the anti-PD-1 treatment group were also given an equivalent volume of normal saline. The drug was administered for 21 days. In the anti-PD-1 antibody group， mice were intraperitoneally injected with 100 μg of mouse anti-PD-1 antibody twice a week， for a total of five injections. The tumor volume， survival time and tumor mass were measured at different time points. Hematoxylin-eosin （HE） staining was used to observe the histological changes of the tumor. The expression of CD8+T cells and TCF1+CD8+ stem-like T cells in tumor tissues was detected by immunohistochemistry and immunofluorescence staining. Flow cytometry was used to detect the difference of immune cell subsets in tumors and the expression difference of TCF1+CD8+ stem cell-like T cells in tumors and peripheral blood. The expression level of PD-L1 in tumor tissues was detected by Western blot. ResultsCompared with model group， the tumor volume and mass of in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group were decreased （P<0.05， P<0.01）. The median survival time of mice in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group was as follows： 27.00 days （95%CI， 0.45-2.65）， 31.00 days （95%CI， 0.32-1.89）， 34.00 days （95%CI， 0.40-2.33）， and 35.00 days （95%CI， 0.42-2.47）. All of them were higher than that of the model group ［24.50 days （95%CI， 0.37-10.5）］. Flow cytometry showed that compared with the model group， the proportion and number of infiltrating CD8+ T cells in tumor were increased in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group （P<0.05， P<0.01）， while the proportion of tumor regulatory T cells （Treg） and M2 macrophages decreased （P<0.05）. Compared with the model group， the proportion of IFN-γ+CD8+ T and GrzB+CD8+ T cells in tumors in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group was increased （P<0.01）， and the proportion of TCF1+CD8+ T cells in tumor and peripheral blood was also increased. Immunofluorescence staining further showed that the number of TCF1+CD8+ T cells in tumor tissues increased in low-， medium- and high-dose Huoluo Xiaolingdan groups. Western blot analysis showed no significant decrease in the PD-L1 protein expression in tumor tissues between the Huoluo Xiaolingdan groups and the model group. ConclusionHuoluo Xiaolingdan can inhibit TNBC in mice by increasing tumor infiltration of TCF1+CD8+ stem-like T cells， enhancing CD8+ T cell activity， and regulating immune cell subgroups such as M2 macrophages and Treg cells to enhance anti-tumor immunity. This study provides a theoretical basis for the clinical application of Huoluo Xiaolingdan in breast cancer treatment and combination therapy.
