Cordycepin Downregulates Cdk-2 to Interfere with Cell Cycle and Increases Apoptosis by Generating ROS in Cervical Cancer Cells: in vitro and in silico Study.
Antineoplastic Agents
/ pharmacology
Apoptosis
/ drug effects
Cell Cycle
/ drug effects
Cell Proliferation
/ drug effects
Computer Simulation
Cyclin-Dependent Kinase 2
/ antagonists & inhibitors
Deoxyadenosines
/ pharmacology
Female
HeLa Cells
Humans
In Vitro Techniques
Reactive Oxygen Species
/ metabolism
Tumor Cells, Cultured
Uterine Cervical Neoplasms
/ drug therapy
Cdk-2.0007U
Cordycepin
apoptosis
cell cycle
cervical cancer
reactive oxygen species.
Journal
Current cancer drug targets
ISSN: 1873-5576
Titre abrégé: Curr Cancer Drug Targets
Pays: Netherlands
ID NLM: 101094211
Informations de publication
Date de publication:
2019
2019
Historique:
received:
08
11
2017
revised:
25
06
2018
accepted:
25
08
2018
pubmed:
6
9
2018
medline:
30
5
2020
entrez:
6
9
2018
Statut:
ppublish
Résumé
Cordycepin is a small molecule from medicinal mushroom Cordyceps, which has been reported for anticancer properties. In this study, we have focused on the investigation of cordycepin effect on cervical cancer cells with further clarification of possible molecular mechanism. We have used cell viability and cell counting assay for cytotoxic effect of cordycepin, flow cytometric assay of apoptosis and cell cycle, and quantitative PCR (qPCR) and Western blotting for the determination of target gene expression. Molecular docking and Molecular dynamics simulation were used for in silico analysis of cordycepin affinity to target protein(s). Treatment of cordycepin controlled SiHa and HeLa cervical cancer cell growth, increased the rate of their apoptosis, and interfered with cell cycle, specifically elongated S-phase. qPCR results indicated that there was a downregulation of cell cycle proteins CDK-2, CYCLIN-A2 and CYCLIN-E1 in mRNA level by cordycepin treatment but no significant change was observed in pro-apoptotic or antiapoptotic proteins. The intracellular reactive oxygen species (ROS) level in cordycepin treated cells was increased significantly, implying that apoptosis might be induced by ROS. Western blot analysis confirmed significant decrease of Cdk-2 and mild decrease of Cyclin-E1 and Cyclin-A2 by cordycepin, which might be responsible for regulating cell cycle. Molecular docking indicated high binding affinity of cordycepin against Cdk-2. Molecular dynamics simulation further confirmed that the docked pose of cordycepin-Cdk-2 complex remained within the binding pocket for 10 ns. Our study suggests that cordycepin is effective against cervical cancer cells, and regulating cell cycle via cell cycle proteins, especially downregulating Cdk-2, and inducing apoptosis by generating ROS are among the mechanisms of anticancer activities of cordycepin.
Sections du résumé
BACKGROUND
Cordycepin is a small molecule from medicinal mushroom Cordyceps, which has been reported for anticancer properties.
OBJECTIVE
In this study, we have focused on the investigation of cordycepin effect on cervical cancer cells with further clarification of possible molecular mechanism.
METHOD
We have used cell viability and cell counting assay for cytotoxic effect of cordycepin, flow cytometric assay of apoptosis and cell cycle, and quantitative PCR (qPCR) and Western blotting for the determination of target gene expression. Molecular docking and Molecular dynamics simulation were used for in silico analysis of cordycepin affinity to target protein(s).
RESULTS
Treatment of cordycepin controlled SiHa and HeLa cervical cancer cell growth, increased the rate of their apoptosis, and interfered with cell cycle, specifically elongated S-phase. qPCR results indicated that there was a downregulation of cell cycle proteins CDK-2, CYCLIN-A2 and CYCLIN-E1 in mRNA level by cordycepin treatment but no significant change was observed in pro-apoptotic or antiapoptotic proteins. The intracellular reactive oxygen species (ROS) level in cordycepin treated cells was increased significantly, implying that apoptosis might be induced by ROS. Western blot analysis confirmed significant decrease of Cdk-2 and mild decrease of Cyclin-E1 and Cyclin-A2 by cordycepin, which might be responsible for regulating cell cycle. Molecular docking indicated high binding affinity of cordycepin against Cdk-2. Molecular dynamics simulation further confirmed that the docked pose of cordycepin-Cdk-2 complex remained within the binding pocket for 10 ns.
CONCLUSION
Our study suggests that cordycepin is effective against cervical cancer cells, and regulating cell cycle via cell cycle proteins, especially downregulating Cdk-2, and inducing apoptosis by generating ROS are among the mechanisms of anticancer activities of cordycepin.
Identifiants
pubmed: 30182857
pii: CCDT-EPUB-92822
doi: 10.2174/1568009618666180905095356
doi:
Substances chimiques
Antineoplastic Agents
0
Deoxyadenosines
0
Reactive Oxygen Species
0
CDK2 protein, human
EC 2.7.11.22
Cyclin-Dependent Kinase 2
EC 2.7.11.22
cordycepin
GZ8VF4M2J8
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
152-159Informations de copyright
Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.