Xi-Ran He1, Shu-Yan Han2, Xiao-Hong Li3, Wen-Xian Zheng4, Li-Na Pang5, Shan-Tong Jiang6, Ping-Ping Li7. 1. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integration of Traditional Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China. Electronic address: hexr2011@126.com. 2. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integration of Traditional Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China. Electronic address: shuyanhan@bjmu.edu.cn. 3. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integration of Traditional Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China. Electronic address: hanxiaolemon@163.com. 4. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integration of Traditional Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China; Department of Oncology, The Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai 200233, China. Electronic address: zhengwenxiansysu@126.com. 5. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integration of Traditional Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China. Electronic address: 18611066280@163.com. 6. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integration of Traditional Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China. Electronic address: wo907152921@126.com. 7. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integration of Traditional Chinese and Western Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China. Electronic address: lppma123@sina.com.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese medicine Bu-Fei decoction (BFD) has been utilized to treat patients with Qi deficiency for decades, with the advantages of invigorating vital energy, clearing heat-toxin and moistening lung, etc. According to previous clinical experience and trials, BFD has been found to indeed improve life quality of lung cancer patients and prolong survival time. Nevertheless, little is known on its potential mechanisms so far. Being regarded as a pivotal cytokine in the tumor microenvironment, transforming growth factor β (TGF-β) stands out as a robust regulator of epithelial-mesenchymal transition (EMT), which is closely linked to tumor progression. AIM OF THE STUDY: The present study was designed to explore whether BFD antagonized EMT via blocking TGF-β1-induced signaling pathway, and then help contribute to create a relatively steady microenvironment for confining lung cancer. MATERIALS AND METHODS: This experiment was performed in lung adenocarcinoma A549 cells both in vitro and in vivo. In detail, the influences mediated by TGF-β1 alone or in combination with different concentrations of BFD on migration were detected by wound healing and transwell assays, and the effects of BFD on cell viability were determined by cell counting kit-8 (CCK-8) assay. TGF-β1, EMT relevant proteins and genes were evaluated by western blotting, confocal microscopy, quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC) and enzyme-linked immuno sorbent assay (ELISA). Female BALB/C nude mice were subcutaneously implanted A549 cells and given BFD by gavage twice daily for 28 days. The tumor volume was monitored every 4 days to draw growth curve. The tumor weight, expression levels of EMT-related protein in tumor tissues and TGF-β1 serum level were evaluated, respectively. RESULTS: BFD only exerted minor effects on A549 cell proliferation and this was in accordance with the in vivo result, which showed that the tumor growth and weight were not be restrained by BFD administration. However, the data elucidated that BFD could dose-dependently suppress EMT induced by TGF-β1 in vitro via attenuating canonical Smad signaling pathway. In the A549 xenograft mouse model, BFD also inhibited protein markers that are associated with EMT and TGF-β1 secretion into serum. CONCLUSIONS: Based on these above data, the conclusion could be put forward that BFD probably attenuated TGF-β1 mediated EMT in A549 cells via decreasing canonical Smad signaling pathway both in vitro and in vivo, which may help restrain the malignant phenotype induced by TGF-β1 in A549 cells to some extent.
ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese medicine Bu-Fei decoction (BFD) has been utilized to treat patients with Qi deficiency for decades, with the advantages of invigorating vital energy, clearing heat-toxin and moistening lung, etc. According to previous clinical experience and trials, BFD has been found to indeed improve life quality of lung cancerpatients and prolong survival time. Nevertheless, little is known on its potential mechanisms so far. Being regarded as a pivotal cytokine in the tumor microenvironment, transforming growth factor β (TGF-β) stands out as a robust regulator of epithelial-mesenchymal transition (EMT), which is closely linked to tumor progression. AIM OF THE STUDY: The present study was designed to explore whether BFD antagonized EMT via blocking TGF-β1-induced signaling pathway, and then help contribute to create a relatively steady microenvironment for confining lung cancer. MATERIALS AND METHODS: This experiment was performed in lung adenocarcinoma A549 cells both in vitro and in vivo. In detail, the influences mediated by TGF-β1 alone or in combination with different concentrations of BFD on migration were detected by wound healing and transwell assays, and the effects of BFD on cell viability were determined by cell counting kit-8 (CCK-8) assay. TGF-β1, EMT relevant proteins and genes were evaluated by western blotting, confocal microscopy, quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC) and enzyme-linked immuno sorbent assay (ELISA). Female BALB/C nude mice were subcutaneously implanted A549 cells and given BFD by gavage twice daily for 28 days. The tumor volume was monitored every 4 days to draw growth curve. The tumor weight, expression levels of EMT-related protein in tumor tissues and TGF-β1 serum level were evaluated, respectively. RESULTS: BFD only exerted minor effects on A549 cell proliferation and this was in accordance with the in vivo result, which showed that the tumor growth and weight were not be restrained by BFD administration. However, the data elucidated that BFD could dose-dependently suppress EMT induced by TGF-β1 in vitro via attenuating canonical Smad signaling pathway. In the A549 xenograft mouse model, BFD also inhibited protein markers that are associated with EMT and TGF-β1 secretion into serum. CONCLUSIONS: Based on these above data, the conclusion could be put forward that BFD probably attenuated TGF-β1 mediated EMT in A549 cells via decreasing canonical Smad signaling pathway both in vitro and in vivo, which may help restrain the malignant phenotype induced by TGF-β1 in A549 cells to some extent.