Ting-Ting Ma1, Gan-Lin Zhang2, Cun-Fang Dai3, Bo-Ran Zhang1, Ke-Xin Cao3, Chun-Guo Wang4, Guo-Wang Yang3, Xiao-Min Wang5. 1. Oncology Department, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, No.23 Back Road of Art Gallery, Dongcheng District, Beijing, 100010, China; Beijing University of Chinese Medicine, No. 11 East North Third Ring Road, Chaoyang District, Beijing, 100029, China. 2. Oncology Department, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, No.23 Back Road of Art Gallery, Dongcheng District, Beijing, 100010, China. Electronic address: kalinezhang@163.com. 3. Oncology Department, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, No.23 Back Road of Art Gallery, Dongcheng District, Beijing, 100010, China. 4. Beijing University of Chinese Medicine, No. 11 East North Third Ring Road, Chaoyang District, Beijing, 100029, China. 5. Oncology Department, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, No.23 Back Road of Art Gallery, Dongcheng District, Beijing, 100010, China. Electronic address: wangxiaomin_bhtcm@126.com.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: The Scutellaria barbata and Hedyotis diffusa (SH) herb pair is extensively used in Traditional Chinese Medicine for efficacy enhancement in cancer treatment in China and Asian countries. Superior clinical efficacy observations based on high dosages (≥60 g) motivated us to explore appropriate dosages and the underlying mechanisms of action. AIM OF THE STUDY: To explore the efficacy and potential mechanisms of actions of SH through in vitro and in vivo experiments and network pharmacology. MATERIALS AND METHODS: SH lyophilized powder (SHLP) was prepared from decoctions and the active ingredients were identified using high performance liquid chromatography (HPLC). Proliferation and migration experiments in vitro and tumor growth in vivo were performed to evaluate the effects of SHLP on breast cancer. Corresponding potential target genes for SHLP components and breast cancer were extracted from established databases and the Protein-Protein Internetwork of shared genes were constructed using STRING database. Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotation clusters were acquired and the top 30 pathways were presented. At last, as one of pathways indicated by enriched results, apoptosis was validated with flow cytometric analysis and caspase-3, 8, 9 activities. RESULTS: Seventy-five ingredients were identified from SHLP by HPLC. High SHLP doses inhibited proliferation and migration of three types of breast cancer cells in vitro and tumor growth in nude mice. After target genes extraction and intersection, the top 30 KEGG clusters were enriched, including PI3K-Akt, cell cycle and other related pathways like VEGF, Micro-RNAs and NF-κB, besides, key genes in apoptosis were mapped. In the last, apoptosis was validated by flow cytometric analysis and caspase-3, 8, 9 activities after SHLP treatment. CONCLUSION: High SHLP dosages inhibited breast cancer in vitro and in vivo, enriched by network pharmacology and confirmed by flow cytometric analysis and caspase activation, with apoptosis was identified as one of the mechanisms of action of SHLP. SHLP administration with higher doses is recommended for clinical usage.
ETHNOPHARMACOLOGICAL RELEVANCE: The Scutellaria barbata and Hedyotis diffusa (SH) herb pair is extensively used in Traditional Chinese Medicine for efficacy enhancement in cancer treatment in China and Asian countries. Superior clinical efficacy observations based on high dosages (≥60 g) motivated us to explore appropriate dosages and the underlying mechanisms of action. AIM OF THE STUDY: To explore the efficacy and potential mechanisms of actions of SH through in vitro and in vivo experiments and network pharmacology. MATERIALS AND METHODS: SH lyophilized powder (SHLP) was prepared from decoctions and the active ingredients were identified using high performance liquid chromatography (HPLC). Proliferation and migration experiments in vitro and tumor growth in vivo were performed to evaluate the effects of SHLP on breast cancer. Corresponding potential target genes for SHLP components and breast cancer were extracted from established databases and the Protein-Protein Internetwork of shared genes were constructed using STRING database. Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotation clusters were acquired and the top 30 pathways were presented. At last, as one of pathways indicated by enriched results, apoptosis was validated with flow cytometric analysis and caspase-3, 8, 9 activities. RESULTS: Seventy-five ingredients were identified from SHLP by HPLC. High SHLP doses inhibited proliferation and migration of three types of breast cancer cells in vitro and tumor growth in nude mice. After target genes extraction and intersection, the top 30 KEGG clusters were enriched, including PI3K-Akt, cell cycle and other related pathways like VEGF, Micro-RNAs and NF-κB, besides, key genes in apoptosis were mapped. In the last, apoptosis was validated by flow cytometric analysis and caspase-3, 8, 9 activities after SHLP treatment. CONCLUSION: High SHLP dosages inhibited breast cancer in vitro and in vivo, enriched by network pharmacology and confirmed by flow cytometric analysis and caspase activation, with apoptosis was identified as one of the mechanisms of action of SHLP. SHLP administration with higher doses is recommended for clinical usage.