Antimalarial Dihydroartemisinin triggers autophagy within HeLa cells of human cervical cancer through Bcl-2 phosphorylation at Ser70.


Journal

Phytomedicine : international journal of phytotherapy and phytopharmacology
ISSN: 1618-095X
Titre abrégé: Phytomedicine
Pays: Germany
ID NLM: 9438794

Informations de publication

Date de publication:
Jan 2019
Historique:
received: 11 06 2018
revised: 25 08 2018
accepted: 25 09 2018
entrez: 3 1 2019
pubmed: 3 1 2019
medline: 17 4 2019
Statut: ppublish

Résumé

As an effective antimalarial medicine, Dihydroartemisinin (DHA) has therapeutic potential on human cervical cancer. However, its working mechanism has not been elucidated. This study aimed to investigate the reversal effect of DHA on human cervical cancer HeLa cells, and explored its mechanism of action in vitro and in vivo. The effect and mechanism of DHA on HeLa cells was examined by using CCK-8 assay, flow cytometry, transmission electron microscopy, immunofluorescence, and Western blot analysis in human hepatocellular carcinoma cells. In this study, it was confirmed that DHA had statistically equivalent anti-tumor efficiency in HeLa cells with a clinical chemotherapeutic agent of cisplatin. Meanwhile, DHA triggered autophagy, where LC3B-II expression was dose-dependently increased. Further, it was revealed that DHA promotes reactive oxygen species (ROS) generation, with DNA double-strand breaks (DSB) damage, as up-regulation of γH2AX protein and foci formation. Interestingly, we firstly demonstrated that DHA induced autophagy through promotion of the phosphorylation of Bcl-2 (Ser70), independent of the phosphorylated JNK1/2 (Thr183/Tyr185). Moreover, DHA-treated HeLa cells displayed an increase in the pro-autophagic protein Beclin-1 with downregulated the phospho-mTOR (Ser2448). Furthermore, upregulated pro-apoptotic protein Bak-1, but not Bax, suggesting Bak-1 is included in DHA-induced autophagy. Therefore, DHA upregulates the phosphorylation of Bcl-2 (Ser70) and mTOR (Ser2448) and induces autophagic cell death in Hela cells. This study provided a mechanism to support DHA, an autophagy inducer, as a potential therapeutic agent for human cervical cancer.

Sections du résumé

BACKGROUND BACKGROUND
As an effective antimalarial medicine, Dihydroartemisinin (DHA) has therapeutic potential on human cervical cancer. However, its working mechanism has not been elucidated.
PURPOSE OBJECTIVE
This study aimed to investigate the reversal effect of DHA on human cervical cancer HeLa cells, and explored its mechanism of action in vitro and in vivo.
STUDY DESIGN/METHODS METHODS
The effect and mechanism of DHA on HeLa cells was examined by using CCK-8 assay, flow cytometry, transmission electron microscopy, immunofluorescence, and Western blot analysis in human hepatocellular carcinoma cells.
RESULTS RESULTS
In this study, it was confirmed that DHA had statistically equivalent anti-tumor efficiency in HeLa cells with a clinical chemotherapeutic agent of cisplatin. Meanwhile, DHA triggered autophagy, where LC3B-II expression was dose-dependently increased. Further, it was revealed that DHA promotes reactive oxygen species (ROS) generation, with DNA double-strand breaks (DSB) damage, as up-regulation of γH2AX protein and foci formation. Interestingly, we firstly demonstrated that DHA induced autophagy through promotion of the phosphorylation of Bcl-2 (Ser70), independent of the phosphorylated JNK1/2 (Thr183/Tyr185). Moreover, DHA-treated HeLa cells displayed an increase in the pro-autophagic protein Beclin-1 with downregulated the phospho-mTOR (Ser2448). Furthermore, upregulated pro-apoptotic protein Bak-1, but not Bax, suggesting Bak-1 is included in DHA-induced autophagy.
CONCLUSION CONCLUSIONS
Therefore, DHA upregulates the phosphorylation of Bcl-2 (Ser70) and mTOR (Ser2448) and induces autophagic cell death in Hela cells. This study provided a mechanism to support DHA, an autophagy inducer, as a potential therapeutic agent for human cervical cancer.

Identifiants

pubmed: 30599894
pii: S0944-7113(18)30503-8
doi: 10.1016/j.phymed.2018.09.221
pii:
doi:

Substances chimiques

Antimalarials 0
Antineoplastic Agents 0
Artemisinins 0
BAK1 protein, human 0
BCL2 protein, human 0
Beclin-1 0
Proto-Oncogene Proteins c-bcl-2 0
Reactive Oxygen Species 0
bcl-2 Homologous Antagonist-Killer Protein 0
artenimol 6A9O50735X
MTOR protein, human EC 2.7.1.1
TOR Serine-Threonine Kinases EC 2.7.11.1
Cisplatin Q20Q21Q62J

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

147-156

Informations de copyright

Copyright © 2018. Published by Elsevier GmbH.

Auteurs

Li Wang (L)

Laboratory of Organ Fibrosis Prophylaxis and Treatment by Combine Traditional Chinese and Western Medicine, Research Center of Combine Traditional Chinese and Western Medicine, Clinical Laboratory, Affiliated Traditional Medicine Hospital of Southwest Medical University, Luzhou 646000, China.

Jianchun Li (J)

Laboratory of Organ Fibrosis Prophylaxis and Treatment by Combine Traditional Chinese and Western Medicine, Research Center of Combine Traditional Chinese and Western Medicine, Clinical Laboratory, Affiliated Traditional Medicine Hospital of Southwest Medical University, Luzhou 646000, China.

Xinli Shi (X)

Hebei Provincial Engineering Laboratory of Plant Bioreactor Preparation Technology, Department of Pathobiology and Immunology, Hebei University of Chinese Medicine, Shijiazhuang, China; Department of Otolaryngology Head and Neck Surgery, Bethune International Peace Hospital, Shijiazhuang 050081, China. Electronic address: sxlsunshine@sina.com.

Shenghao Li (S)

Hebei Provincial Engineering Laboratory of Plant Bioreactor Preparation Technology, Department of Pathobiology and Immunology, Hebei University of Chinese Medicine, Shijiazhuang, China.

Patrick Ming-Kuen Tang (PM)

Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, Hong Kong 999077, China.

Zhen Li (Z)

Department of Otolaryngology Head and Neck Surgery, Bethune International Peace Hospital, Shijiazhuang 050081, China.

Hui Li (H)

Department of Otolaryngology Head and Neck Surgery, Bethune International Peace Hospital, Shijiazhuang 050081, China.

Cong Wei (C)

Laboratory of Organ Fibrosis Prophylaxis and Treatment by Combine Traditional Chinese and Western Medicine, Research Center of Combine Traditional Chinese and Western Medicine, Clinical Laboratory, Affiliated Traditional Medicine Hospital of Southwest Medical University, Luzhou 646000, China.

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Classifications MeSH