Li Qiong, Chen Jing-Xian, Wu Yuan, Lv Ling-Ling, Ying Hai-Feng, Zhu Wen-Hua, Xu Jia-Yue, Ruan Ming, Guo Yuanbiao, Zhu Wei-Rong, Zheng Lan
Department of Traditional Chinese Medicine, Shanghai Jiao Tong University School of Medicine Affiliated Ruijin Hospital, Shanghai, 200025, China.
J Ethnopharmacol. 2022 Apr 6;287:114904. doi: 10.1016/j.jep.2021.114904. Epub 2021 Dec 21.
Fuzheng Xiaojijinzhan (FZXJJZF) decoction is an effective prescription for treating colorectal cancer liver metastasis (LMCRC).
To elucidate the pharmacological mechanism of the FZXJJZF decoction therapy on LMCRC.
Firstly, a network pharmacological approach was used to characterize the underlying targets of FZXJJZF on LMCRC. Secondly, LMCRC-related genes are obtained from the public database TCGA, and those genes are further screened and clustered through Mfuzz, an R package tool. Then, targets of FZXJJZF predicted by network pharmacology were overlapped with LMCRC related genes screened by Mfuzz. Meanwhile, FZJZXJF intervened in LMCRC model,epithelial-to-mesenchymal transition (EMT), and migration and invasion of HCT-116 cells. Thirdly, the transcriptomics data of FZJZXJF inhibited HCT-116 cells of EMT cells were overlapped with EMT database data to narrow the possible range of targets. Based on this, the potential targets and signal pathways of FZJZXJF were speculated by combining the transcriptomics data with the targets from network pharmacology-TCGA. Finally, the anti-cancer mechanism of FZXJJZF on LMCRC was verified in vitro by Real-Time PCR and Western Blot in vitro.
By network pharmacological analysis, 282 ingredients and 429 potential targets of FZXJJZF were predicted. The 9268 LMCRC-related genes in the TCGA database were classified into 10 clusters by the Mfuzz. The two clustering genes with the most similar clustering trends were overlapped with 429 potential targets, and 32 genes were found, such as CD34, TRPV3, PGR, VDR, etc. In vivo experiments, FZJZXJF inhibited the tumor size in LMCRC models, and the EMT, migration, and invasion of HCT-116 also be inhibited. Intersecting transcriptomics dates with 32 target genes, it is speculated that the VDR-TGF-β signaling pathway may be an effective mechanism of FZXJJZF. Additionally, it is shown that FZXJJZF up-regulated the expression levels of VDR and E-cadherin and down-regulated the expression levels of TGF-β and Snail1 in vitro. These results confirmed that FZXJJZF plays an effective role in LMCRC mainly by inhibiting EMT phenotype via the VDR-TGF-β signaling pathway.
Collectively, this study reveals the anti-LMCRC effect of FZXJJZF and its potential therapeutic mechanism from the perspective of potential targets and potential pathways.
扶正消积软坚方（FZXJJZF）汤剂是治疗结直肠癌肝转移（LMCRC）的有效方剂。
阐明FZXJJZF汤剂治疗LMCRC的药理机制。
首先，采用网络药理学方法表征FZXJJZF对LMCRC的潜在靶点。其次，从公共数据库TCGA中获取与LMCRC相关的基因，并通过R包工具Mfuzz对这些基因进行进一步筛选和聚类。然后，将网络药理学预测的FZXJJZF靶点与Mfuzz筛选的LMCRC相关基因进行重叠。同时，FZJZXJF干预LMCRC模型、上皮-间质转化（EMT）以及HCT-116细胞的迁移和侵袭。第三，将FZJZXJF抑制EMT细胞的HCT-116细胞的转录组学数据与EMT数据库数据进行重叠，以缩小可能的靶点范围。基于此，结合转录组学数据与网络药理学-TCGA的靶点，推测FZJZXJF的潜在靶点和信号通路。最后，通过实时荧光定量PCR和蛋白质免疫印迹在体外验证FZXJJZF对LMCRC的抗癌机制。
通过网络药理学分析，预测出FZXJJZF的282种成分和429个潜在靶点。TCGA数据库中9268个与LMCRC相关的基因通过Mfuzz被分为10个簇。将聚类趋势最相似的两个聚类基因与429个潜在靶点进行重叠，发现了32个基因，如CD34、TRPV3、PGR、VDR等。体内实验中，FZJZXJF抑制了LMCRC模型中的肿瘤大小，HCT-116的EMT、迁移和侵袭也受到抑制。将转录组学数据与32个靶基因进行交叉分析，推测VDR-TGF-β信号通路可能是FZXJJZF的有效作用机制。此外，体外实验表明FZXJJZF上调了VDR和E-钙黏蛋白的表达水平，下调了TGF-β和Snail1的表达水平。这些结果证实FZXJJZF在LMCRC中主要通过VDR-TGF-β信号通路抑制EMT表型发挥有效作用。
总体而言，本研究从潜在靶点和潜在通路的角度揭示了FZXJJZF对LMCRC的抗癌作用及其潜在治疗机制。
