Digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathway

Abstract Hepatocellular carcinoma (HCC) is recognized as a highly malignant tumor. Targeted combination immunotherapy, the initially approved regimen, is compromised by adverse side effects and low response rates during clinical treatment. Traditional Chinese medicine and its derived natural compoun...

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Main Authors: Mengqing Ma, Rui Hu, Qi Huang, Jing Li, Minling Lv, Jialing Sun, Xin Zhong, Jinyu Yi, Lanfen Peng, Wenxing Feng, Wenfeng Ma, Zhiyi Han, Wei Zhang, Xinfeng Sun, Bolin Zhan, Xingning Liu, Xiaozhou Zhou
Format: Article
Language:English
Published: BMC 2024-12-01
Series:Cancer Cell International
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Online Access:https://doi.org/10.1186/s12935-024-03602-z
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author Mengqing Ma
Rui Hu
Qi Huang
Jing Li
Minling Lv
Jialing Sun
Xin Zhong
Jinyu Yi
Lanfen Peng
Wenxing Feng
Wenfeng Ma
Zhiyi Han
Wei Zhang
Xinfeng Sun
Bolin Zhan
Xingning Liu
Xiaozhou Zhou
author_facet Mengqing Ma
Rui Hu
Qi Huang
Jing Li
Minling Lv
Jialing Sun
Xin Zhong
Jinyu Yi
Lanfen Peng
Wenxing Feng
Wenfeng Ma
Zhiyi Han
Wei Zhang
Xinfeng Sun
Bolin Zhan
Xingning Liu
Xiaozhou Zhou
author_sort Mengqing Ma
collection DOAJ
description Abstract Hepatocellular carcinoma (HCC) is recognized as a highly malignant tumor. Targeted combination immunotherapy, the initially approved regimen, is compromised by adverse side effects and low response rates during clinical treatment. Traditional Chinese medicine and its derived natural compounds, known for their anticancer effects, offer advantages of low toxicity and cost. In this study, we performed high-throughput phenotypic screening in vitro to identify promising anti-HCC drugs. Among 1,444 bioactive compounds, digoxigenin (DIG) was found to significantly impede HCC cell progression. We validated DIG’s therapeutic effects through assays such as cell counting by CCK8, lactate dehydrogenase, and colony formation. Analyses including transmission electron microscopy, western blotting, and immunofluorescence demonstrated that DIG inhibits HCC cell proliferation via autophagy. Network pharmacology and molecular docking studies suggest that DIG targets the PI3K/AKT/mTOR signaling pathway. Comparative treatments of Hep3B and Huh7 cells with DIG or mTOR inhibitors revealed similar inhibitory impacts, indicating that DIG induces autophagy by inhibiting the PI3K/AKT/mTOR pathway. In vivo studies confirmed that DIG halts the growth of subcutaneous xenograft tumors. In conclusion, DIG represents a potential HCC treatment by modulating the PI3K/AKT/mTOR pathway to induce autophagy. This research, via phenotypic screening, accelerates drug discovery and the development of novel therapies targeting the underlying mechanisms of liver cancer.
format Article
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institution Kabale University
issn 1475-2867
language English
publishDate 2024-12-01
publisher BMC
record_format Article
series Cancer Cell International
spelling doaj-art-0021a90d0a6b424c9c988c053f8c03e92024-12-22T12:49:17ZengBMCCancer Cell International1475-28672024-12-0124111710.1186/s12935-024-03602-zDigoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathwayMengqing Ma0Rui Hu1Qi Huang2Jing Li3Minling Lv4Jialing Sun5Xin Zhong6Jinyu Yi7Lanfen Peng8Wenxing Feng9Wenfeng Ma10Zhiyi Han11Wei Zhang12Xinfeng Sun13Bolin Zhan14Xingning Liu15Xiaozhou Zhou16Department of Liver Disease, Shenzhen Traditional Chinese Medicine HospitalDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, Shenzhen Traditional Chinese Medicine HospitalDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineFaculty of Chinese Medicine, Macau University of Science and TechnologyDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineDepartment of Liver Disease, The Fourth Clinical Medical College of Guangzhou University of Chinese MedicineAbstract Hepatocellular carcinoma (HCC) is recognized as a highly malignant tumor. Targeted combination immunotherapy, the initially approved regimen, is compromised by adverse side effects and low response rates during clinical treatment. Traditional Chinese medicine and its derived natural compounds, known for their anticancer effects, offer advantages of low toxicity and cost. In this study, we performed high-throughput phenotypic screening in vitro to identify promising anti-HCC drugs. Among 1,444 bioactive compounds, digoxigenin (DIG) was found to significantly impede HCC cell progression. We validated DIG’s therapeutic effects through assays such as cell counting by CCK8, lactate dehydrogenase, and colony formation. Analyses including transmission electron microscopy, western blotting, and immunofluorescence demonstrated that DIG inhibits HCC cell proliferation via autophagy. Network pharmacology and molecular docking studies suggest that DIG targets the PI3K/AKT/mTOR signaling pathway. Comparative treatments of Hep3B and Huh7 cells with DIG or mTOR inhibitors revealed similar inhibitory impacts, indicating that DIG induces autophagy by inhibiting the PI3K/AKT/mTOR pathway. In vivo studies confirmed that DIG halts the growth of subcutaneous xenograft tumors. In conclusion, DIG represents a potential HCC treatment by modulating the PI3K/AKT/mTOR pathway to induce autophagy. This research, via phenotypic screening, accelerates drug discovery and the development of novel therapies targeting the underlying mechanisms of liver cancer.https://doi.org/10.1186/s12935-024-03602-zHepatocellular carcinomaDIGAutophagyPI3K/AKT/mTOR pathwaySmall-molecule compounds
spellingShingle Mengqing Ma
Rui Hu
Qi Huang
Jing Li
Minling Lv
Jialing Sun
Xin Zhong
Jinyu Yi
Lanfen Peng
Wenxing Feng
Wenfeng Ma
Zhiyi Han
Wei Zhang
Xinfeng Sun
Bolin Zhan
Xingning Liu
Xiaozhou Zhou
Digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathway
Cancer Cell International
Hepatocellular carcinoma
DIG
Autophagy
PI3K/AKT/mTOR pathway
Small-molecule compounds
title Digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathway
title_full Digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathway
title_fullStr Digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathway
title_full_unstemmed Digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathway
title_short Digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathway
title_sort digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the pi3k akt mtor pathway
topic Hepatocellular carcinoma
DIG
Autophagy
PI3K/AKT/mTOR pathway
Small-molecule compounds
url https://doi.org/10.1186/s12935-024-03602-z
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