Affiliation:
1. Jiangxi University of Chinese Medicine, Nanchang 330004, China
2. Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic
Drug Development, Nanning 530200, China
3. College of Pharmacy, Guangxi University of Chinese
Medicine, Nanning 530000, China
4. Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning,
330004, China
Abstract
Background:
Necroptosis is a type of programmed necrosis mediated by receptor-interacting
protein kinases 1 and 3 (RIP1 and RIP3), which is morphologically characterized by enlarged
organelles, ruptured plasma membrane, and subsequent loss of intracellular contents. Cryptotanshinone
(CPT), a diterpene quinone compound extracted from the root of Salvia miltiorrhiza
Bunge, has been reported to have significant anticancer activities. However, the detailed mechanism
of CPT has not been clearly illustrated.
Objective:
The present study aimed to explore the cell death type and mechanisms of CPT-induced
in non-small cell lung cancer (NSCLC) cells.
Methods:
The cytotoxicity of CPT on A549 cells was assessed by MTS assay. Ca2+ release and reactive
oxygen species (ROS) generation were detected by flow cytometry. The changes in mitochondrial
membrane potential (MMP) were observed through JC-1 staining. The expressions of p-
RIP1, p-RIP3, p-MLKL, and MAPKs pathway proteins were analyzed by western blotting analysis.
The efficacy of CPT in vivo was evaluated by the Lewis lung carcinoma (LLC) xenograft mice
model. Blood samples were collected for hematology analysis. ELISA investigated the effects of
CPT on tumor necrosis factor α (TNF-α). Hematoxylin and eosin staining (HE) determined the tumor
tissues. Proteins' expression of tumor tissues was quantified by western blotting.
Results:
CPT inhibited the cell viability of A549 cells in a time- and concentration-dependent manner,
which was reversed by Necrostatin-1 (Nec-1). In addition, CPT treatment increased the expression
of p-RIP1, p-RIP3, p-MLKL, the release of Ca2+, ROS generation, and the MAPKs pathway
activated in A549 cells. Moreover, animal experiment results showed that intraperitoneal injection
of CPT (15 mg/kg and 30 mg/kg) significantly inhibited tumor growth in C57BL/6 mice without affecting
the bodyweight and injuring the organs.
Conclusion:
Our findings suggested that CPT-induced necroptosis via RIP1/RIP3/MLKL signaling
pathway both in vitro and in vivo, indicating that CPT may be a promising agent in the treatment
of NSCLC.
Funder
Guangxi University of Chinese Medicine
Guangxi Key Research and Development Plan
Guangxi Science and Technology Base and Talent Special Project
National Natural Science Foundation of China
Guangxi Natural Science Foundation
Publisher
Bentham Science Publishers Ltd.
Subject
General Health Professions
Cited by
2 articles.
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