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Proteomic study on sodium selenite-induced apoptosis of human cervical cancer HeLa cells
Liping Fu1, Qiong Liu, Liming Shen
1College of Life Sciences, Shenzhen University, Shenzhen 518060, China.
Abstract:
Sodium selenite can induce the apoptosis of cancer cells, however its mechanism has seldom been studied via proteomics. In this paper, human cervical cancer HeLa cells were investigated by MTT assay and morphological observation to get appropriate selenite concentrations for proteomic study. Results showed that selenite at concentrations larger than 10 μmol/L significantly inhibited the viability of HeLa cells. 40 μmol/L selenite was in the appropriate range for proteomic study. After 24 h treatment with 40 μmol/L selenite, total proteins were extracted from the cells and applied to two-dimensional gel electrophoresis (2DE). Those proteins with their expression levels altered at least 2-fold comparing to the control were picked up for protein identification via MALDI-TOF mass spectrometry and further confirmed by Western blot analysis. About 1000 spots were detected by the software in each 2DE gel, among which 13 differentially expressed proteins were identified by mass spectrometry and most of them are relevant to oxidative stress, such as peroxiredoxins, superoxide dismutase, quinolinate phosphoribosyl transferase, and D-dopachrome tautomerase. Meanwhile, reactive oxygen species (ROS) and mitochondrial membrane potential were also detected by flow cytometry and laser confocal scanning microscope. An increase in ROS generation and a decrease in mitochondrial membrane potential were detected in the selenite-treated cells compared with the control, which are consistent with the down-expression of antioxidative proteins in proteomics. Those results indicate that selenite induces the apoptosis of HeLa cells via ROS-mediated mitochondrial pathway. The present study also implies the potentiality of selenium in cervical cancer treatment.
Insights
Sodium selenite induces cancer cell death by increasing oxidative stress and damaging mitochondria. This study used proteomics to identify proteins involved in selenite-induced apoptosis in HeLa cells, revealing potential for selenium in cervical cancer treatment.
Area of Science:
- Biochemistry
- Cell Biology
- Oncology
Background:
- Sodium selenite demonstrates anticancer properties by inducing apoptosis.
- The precise molecular mechanisms, particularly via proteomics, remain under-investigated.
- Cervical cancer remains a significant global health concern requiring novel therapeutic strategies.
Purpose of the Study:
- To investigate the proteomic changes in human cervical cancer HeLa cells treated with sodium selenite.
- To elucidate the mechanism of selenite-induced apoptosis in HeLa cells.
- To identify potential therapeutic targets for cervical cancer treatment using selenium.
Main Methods:
- Cell viability was assessed using MTT assay and morphological observation.
- Proteomic analysis involved two-dimensional gel electrophoresis (2DE), MALDI-TOF mass spectrometry, and Western blot.
- Reactive oxygen species (ROS) generation and mitochondrial membrane potential were measured using flow cytometry and laser confocal microscopy.
Main Results:
- Sodium selenite at concentrations above 10 μmol/L significantly inhibited HeLa cell viability, with 40 μmol/L identified as optimal for proteomic study.
- Proteomic analysis identified 13 differentially expressed proteins, many associated with oxidative stress, including peroxiredoxins and superoxide dismutase.
- Increased ROS generation and decreased mitochondrial membrane potential were observed in selenite-treated cells, correlating with the downregulation of antioxidative proteins.
Conclusions:
- Sodium selenite induces apoptosis in HeLa cells through a reactive oxygen species (ROS)-mediated mitochondrial pathway.
- The identified differentially expressed proteins provide insights into the cellular response to selenite treatment.
- Selenium compounds show promise as a potential therapeutic agent for cervical cancer.
