Related Experiment Video
Updated: Dec 6, 2025
A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage
Published on: May 10, 2022
Topotecan induces hepatocellular injury via ASCT2 mediated oxidative stress
Guoliang Zhou1, Meisong Qin1, Xiaolin Zhang1
1Department of Pharmacy, School of Life and Health Sciences, Anhui Science and Technology University, Fengyang, Anhui, China.
Background:
Topotecan is an anti-cancer chemotherapy drug with common side effects, including hepatotoxicity. In this study, we aim to investigate the mechanisms of topotecan-induced hepatocellular injury beyond conventional DNA damage.
Materials And Methods:
Methyl Thiazolyl Tetrazolium (MTT) assay was used to detect the inhibitory effect of topotecan on cell proliferation. Western blot was used to detect protein expression. Flow cytometry assay was performed to determine apoptosis rate under topotecan treatment. ASCT2 overexpression was addressed using adenovirus vector. qRT-PCR and western blot assay were used to detect the expression of ASCT2. Glutamine uptake, intracellular glutathione (GSH) and reactive oxygen species (ROS) level were detected by glutamine detection kit, GSH detection kit and ROS detection kit respectively.
Results:
MTT results showed that topotecan had an inhibitory effect on cell proliferation and induced apoptosis in both L02 and HepG2 cell lines. Topotecan inhibited the expression of glutamine transporter ASCT2 and the uptake of glutamine in both L02 and HepG2 cell lines. The uptake of glutamine and the GSH level was increased in both L02 and HepG2 cell lines after ASCT2 overexpression. The ROS level was inhibited by ASCT2 overexpression upon topotecan treatment in both L02 and HepG2 cell lines. Topotecan-induced hepatocellular apoptosis and proliferation inhibition were attenuated by ASCT2 overexpression in both L02 and HepG2 cell lines.
Conclusion:
Topotecan-induced hepatocytes death is dependent on ASCT2 down-regulation, which causes oxidative stress via inhibiting GSH production.
Insights
Topotecan chemotherapy causes liver injury by downregulating the ASCT2 transporter, reducing glutathione and increasing oxidative stress. Restoring ASCT2 levels protects against topotecan-induced hepatocellular damage.
Area of Science:
- Hepatology
- Oncology
- Biochemistry
Background:
- Topotecan is an anti-cancer drug known to cause hepatotoxicity.
- Mechanisms of topotecan-induced liver injury beyond DNA damage require elucidation.
Purpose of the Study:
- Investigate the role of ASCT2 in topotecan-induced hepatocellular injury.
- Determine the impact of ASCT2 modulation on cell proliferation, apoptosis, oxidative stress, and glutathione levels.
Main Methods:
- Cell proliferation assays (MTT) and apoptosis assays (flow cytometry) were employed.
- Western blot and qRT-PCR assessed ASCT2 expression.
- Glutamine uptake, glutathione (GSH), and reactive oxygen species (ROS) levels were quantified.
Main Results:
- Topotecan inhibited proliferation and induced apoptosis in L02 and HepG2 cells.
- Topotecan decreased ASCT2 expression and glutamine uptake.
- ASCT2 overexpression increased glutamine uptake and GSH, reduced ROS, and attenuated topotecan-induced injury.
Conclusions:
- Topotecan-induced hepatocyte death is linked to ASCT2 downregulation.
- ASCT2 downregulation contributes to oxidative stress by inhibiting GSH production.
More Related Videos
19:44Enhancement of Apoptotic and Autophagic Induction by a Novel Synthetic C-1 Analogue of 7-deoxypancratistatin in Human Breast Adenocarcinoma and Neuroblastoma Cells with Tamoxifen
Published on: May 30, 2012
09:32Drug-Induced Senescence in Liver Cells Promotes M2 Macrophage Polarization: Implications for Tyrosine Kinase Inhibitor-Associated Hepatotoxicity
Published on: October 17, 2025
Related Concept Videos
01:46Electron Transport Chain: Complex I and II
ROS generation is regulated and maintained at moderate levels necessary...
01:19Allosteric Proteins-ATCase
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
01:25Mutagenicity and Carcinogenicity