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Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model
Published on: April 16, 2014
Lactonic sophorolipid-induced apoptosis in human HepG2 cells through the Caspase-3 pathway
Xiao Wang1,2, Na Xu3,4, Qinglin Li1
1Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui University of Chinese Medicine, Hefei, 230038, Anhui, China.
Abstract:
Liver cancer, one of the most common types of cancer in the world, is the second leading cause of death for cancer patients. For liver cancer, there is an urgent need for an effective treatment with no or less toxic side effects. Lactonic sophorolipids (LSL), as a potential anticancer drug, has attracted wide attention of pharmaceutical researchers with its good biological activities. The effects of LSL and cell death inhibitors were measured by MTT test on HepG2 cells. Meanwhile, the morphology of the cells was observed under a microscope. The apoptosis rate was detected by flow cytometry, and the expression levels of enzyme activity of Caspase-3 and Caspase-9 were measured by detection kits. Meanwhile, mRNA levels of Apaf-1, Caspase-3, Bax, and Bcl-2 were measured by quantitative real-time RT-PCR; protein levels of Caspase-3, Cleaved Caspase-3, Bax, and Bcl-2 were measured by western blot. LSL can inhibit the proliferation of cells, and it is possible to induce apoptosis in cells. The HepG2 cells with LSL co-culture exhibited typical apoptotic morphology, and the expression levels of enzyme activity of Caspase-3 and Caspase-9 increased (P< 0.05). We also found that LSL increases cell apoptosis rate and regulates the expression of genes and proteins associated with apoptosis through the Caspase-3 pathway. These results indicate that LSL may be one of the potential drug candidates to inhibit the proliferation and induce apoptosis in HepG2 cells.Key points• LSL, which is of good biological activities such as anti-bacterium, virus elimination, and inflammatory response elimination, has been firstly used to intervene in vitro to investigate its effect on HepG2 cell proliferation.• LSL can inhibit the proliferation of cells, and it is possible to induce apoptosis in HepG2 cells through the Caspase-3 pathway.• The mechanism of LSL action on HepG2 cell proliferation was firstly also discussed, which provides a certain experimental reference for the clinical treatment of liver cancer.
Insights
Lactonic sophorolipids (LSL) show potential as a liver cancer treatment by inhibiting HepG2 cell proliferation and inducing apoptosis. LSL activates the Caspase-3 pathway, offering a promising therapeutic strategy with fewer side effects.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Liver cancer is a leading cause of cancer-related mortality worldwide, necessitating novel therapeutic agents with reduced toxicity.
- Lactonic sophorolipids (LSL) possess known biological activities and are being investigated for their anticancer potential.
Purpose of the Study:
- To investigate the in vitro effects of Lactonic sophorolipids (LSL) on HepG2 liver cancer cell proliferation.
- To elucidate the mechanism by which LSL affects HepG2 cells, focusing on apoptosis induction and related pathways.
Main Methods:
- Cell proliferation was assessed using MTT assays on HepG2 cells treated with LSL.
- Apoptosis was evaluated through morphological observation, flow cytometry, and analysis of Caspase-3 and Caspase-9 activity.
- Gene and protein expression related to apoptosis (Apaf-1, Caspase-3, Bax, Bcl-2) were measured via RT-PCR and Western blot.
Main Results:
- Lactonic sophorolipids (LSL) significantly inhibited HepG2 cell proliferation.
- LSL induced typical apoptotic morphology and increased the apoptosis rate in HepG2 cells.
- LSL treatment led to elevated activity of Caspase-3 and Caspase-9, and modulated the expression of apoptosis-related genes and proteins via the Caspase-3 pathway.
Conclusions:
- Lactonic sophorolipids (LSL) demonstrate significant potential in inhibiting liver cancer cell proliferation and inducing apoptosis.
- The Caspase-3 pathway is implicated in the mechanism of LSL-induced apoptosis in HepG2 cells.
- LSL represents a promising candidate for further development as a therapeutic agent for liver cancer.
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