Liuwei Dihuang Erzhiwan Combination Regulate Myeloid-derived Suppressor Cells to Inhibit Breast Cancer Lung Metastasis
10.13422/j.cnki.syfjx.20240641
- VernacularTitle:六味地黄二至丸合剂影响髓源性抑制细胞与乳腺癌肺转移的相关性
- Author:
Lixiang ZHENG
1
;
Zifeng GUO
1
;
Huiwen GUO
1
;
Xiaomin WANG
1
;
Chuanming XU
1
;
Yuliang HU
2
Author Information
1. Jiangxi University of Chinese Medicine， Nanchang 330004， China
2. Xinjian District People's Hospital， Nanchang 330100， China
- Publication Type:Journal Article
- Keywords:
Liuwei Dihuang Erzhiwan combination;
breast cancer;
lung metastasis;
myeloid-derived suppressor cells （MDSCs）;
epithelial-mesenchymal transition （EMT）-related protein expression;
CC motif chemokine 9 （CCL9）/CC motif chemokine receptor 1 （CCR1） expression
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(20):37-45
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the mechanism by which Liuwei Dihuang Erzhiwan combination inhibit the lung metastasis of spontaneous breast cancer in mice by regulating the recruitment of myeloid-derived suppressor cells （MDSCs）. MethodThree hundred and eighty SPF-grade 10-month-old female breeders of Kunming mouse were palpated at the mammary gland site once every 3 days. Mice that have not had a lump touched after being raised for 6 months are used as control group. After tumor development， the mice were randomized into model， positive control （paclitaxel， intraperitoneal injection at 0.01 g·kg-1 every other day for 22 d）， Liuwei Dihuangwan （0.65 g·kg-1·d-1 by gavage）， Erzhiwan （5.41 g·kg-1·d-1 by gavage）， and Liuwei Dihuang Erzhiwan combination （6.05 g·kg-1·d-1 by gavage） groups. The mice were euthanised when the tumor reached a diameter of about 15 mm， and the tumor and lung tissues were collected. The survival time， tumor mass， and lung metastasis rate of tumor-bearing mice were recorded. Hematoxylin-eosin （HE） staining was used to observe the histopathological and morphological changes of mouse tumor and lung tissues. Immunofluorescence （IF） was used to detect the distribution of MDSCs in tissues of mice in each group by double-staining of MDSCs cells with lymphocyte antigen 6 complex site G6D （Ly6G） and CD11 antigen-like family member B （CD11b）. Western blot was employed to determine the protein levels of matrix metalloproteinase-9 （MMP-9）， transforming growth factor-β （TGF-β）， zinc finger transcription factor 1 （Snail1）， and E-cadherin in the tumor tissue and CC motif chemokine 9 （CCL9） and CC motif chemokine receptor 1 （CCR1） in the lung tissue. ResultDuring the modelling period， the paclitaxel group and Chinese medicine intervention groups had longer median number of days of survival and lower tumor weight， lung metastasis rate， and lung nodule than the model group （P<0.05， P<0.01）. HE staining showed an increase in tumor cell necrosis in the paclitaxel group and the Liuwei Dihuang Erzhiwan combination group. The paclitaxel group and Chinese medicine intervention groups had lower fluorescence intensity of MDSCs in the tumor tissue than the model group （P<0.05， P<0.01）. Compared with the normal control group， the model group showed increased fluorescence intensity of MDSCs in the metastatic lung tissue （P<0.01）， which， however， was decreased in the paclitaxel group and Chinese medicine intervention groups （P<0.01）. The model group showed higher protein levels of MMP-9， TGF-β， and Snail1 and lower protein level of E-cadherin in the tumor tissue than in the normal control group （P<0.01）. Compared with model group， paclitaxel and Chinese medicine interventions downregulated the protein levels of MMP-9， TGF-β， and Snail1 （P<0.05， P<0.01） and upregulated the protein level of E-cadherin in the tumor tissue （P<0.01）. Moreover， the Liuwei Dihuang Erzhiwan combination group had lower protein levels of TGF-β and Snail1 than the Liuwei Dihuangwan group and Erzhiwan group （P<0.05）. In the metastatic lung tissue， the expression of CCL9 and CCR1 was higher in the model group than in the normal control group， paclitaxel group， and Chinese medicine intervention groups （P<0.05， P<0.01）. ConclusionLiuwei Dihuang Erzhiwan combination inhibit tumor growth， prolong survival time， and reduce the occurrence of lung metastasis in the mouse model of spontaneous breast cancer by reducing the recruitment of MDSCs in the tumor and lung tissues and modulating the phenotypes of epithelial-mesenchymal transition （EMT）-related molecules and the expression of CCL9/CCR1.
