疏肝益肾方对乳腺癌肺转移微环境基因表达谱及免疫相关基因表达的影响

Effect of Shugan Yishen Recipe on Microenvironment Gene Expression Profile and Immune-Related Gene Expression in Lung Metastasis of Breast Cancer

    摘要
  • 摘要:
    目的 通过研究疏肝益肾方对乳腺癌小鼠肺部微环境基因表达的影响，探讨疏肝益肾方抑制乳腺癌肺转移的作用机制。
    方法 构建乳腺癌原位移植瘤高转移模型，随机分为生理盐水组，疏肝益肾低、中、高(0.5、1、2 g·kg-1)剂量以及多西他赛组(5 mg·kg-1)，干预28 d。HE染色观察肺组织结构情况；对转移灶进行转录组分析，通过生物信息学分析基因表达谱的GO和KEGG通路特点，筛选关键差异基因表达情况，采用qPCR和Western blot分析关键免疫基因的表达。流式细胞术检测髓源抑制细胞(Myeloid suppressor cells，MDSCs)表达情况；ELISA法检测细胞因子和趋化因子Cxcl2、GM-CSF的表达。
    结果 与生理盐水组相比，疏肝益肾方中、高剂量组与多西他赛组肺组织转移结节数量均有相应减少；HE染色提示肺部病理程度有所改善。疏肝益肾方高剂量组与生理盐水组的肺组织微环境有814个差异表达基因，其中713个基因下调，101个基因上调；筛选疏肝益肾组中关键炎性介质基因Nfkbiz、Tnfaip3、Maff、Hspa1a、Hspb1、Cxcl2，与生理盐水组相比，疏肝益肾方组炎性基因显著下调(P < 0.05，P < 0.01)，最终其结果与转录组结果趋势相符。采用Western blot在蛋白质层面进行验证，发现与生理盐水组相比，疏肝益肾方组Nfkbiz、Tnfaip3、Maff、Hspa1a和Cxcl2蛋白表达水平下降(P < 0.05，P < 0.01)，但Hspb1蛋白表达上调(P < 0.01)。疏肝益肾方组和多西他赛组能够抑制MDSCs表达(P < 0.001)，一定程度降低Cxcl2、GM-CSF的表达(P < 0.05，P < 0.01)。
    结论 疏肝益肾方能抑制乳腺癌肺转移，并且调控肺部免疫微环境基因，其调控范围较广。

     

    Abstract:
    Objective To investigate the mechanism of inhibition of breast cancer lung metastasis by Shugan Yishen Recipe by studying the effects of Shugan Yishen Recipe(SGYSR) on gene expression in the lung microenvironment of mice with breast cancer.
    Methods A high metastasis model of breast cancer was constructed, and the mice were randomly divided into saline group, low, medium, high(0.5, 1, 2 g ·kg-1) Shugan Yishen Recipe groups, and docetaxel group (5 mg ·kg-1), and intervened for 28 days. HE staining was used to observe the lung tissue structure; transcriptome analysis was performed on the metastatic foci, and the key differential gene expression was screened by bioinformatics analysis of the GO and KEGG pathway characteristics of the gene expression profiles, and the expression of key immune genes was analyzed by qPCR and Western blot. The expression of MDSCs was detected by flow cytometry; the expression of cytokines and chemokines including Cxcl2, and GM-CSF was detected by ELISA.
    RESULTS Compared with the saline group, the number of metastatic nodules in lung tissues of the middle and high dose groups of Shugan Yishen Recipe and docetaxel group were correspondingly lower; HE staining suggested that the degree of lung pathology was improved. There were 814 differentially expressed genes in the lung microenvironment of the high-dose group of Shugan Yishen Recipe and the saline group, of which 713 genes were down-regulated and 101 genes were up-regulated; screening for key inflammatory mediator genes in the Shugan Yishen Recipe group, Nfkbiz, Tnfaip3, Maff, Hspa1a, Hspb1 and Cxcl2, the inflammatory genes in the Shugan Yishen Recipe, as compared with that of the saline group, were significantly down-regulated (P < 0.05, P < 0.01), and ultimately its results were consistent with the trend of the transcriptome results. Western blot was used to verify the results at the protein level, and it was found that compared with the saline group, the protein expression levels of Nfkbiz, Tnfaip3, Maff, Hspa1a and Cxcl2 in the Shugan Yishen Recipe group were decreased(P < 0.05, P < 0.01), while the protein expression of Hspb1 was up-regulated(P < 0.01). Both the Shugan Yishen Recipe group and the docetaxel group could inhibit the expression of MDSCs (P < 0.001), and the expression of Cxcl2, and GM-CSF decreased to a certain extent (P < 0.05, P < 0.01).
    CONCLUSION Shugan Yishen Recipe can inhibit breast cancer lung metastasis and regulate the lung immune microenvironment genes in a wide range.

     

    • HTML全文
    • 参考文献(32)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回