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Triterpenoid pristimerin induced HepG2 cells apoptosis through ROS-mediated mitochondrial dysfunction
Yan Guo1, Wei Zhang, Yan-Yan Yan
1Wuhai Municipal People's Hospital, Wuhai, Inner Mongolia, People's Republic of China.
Purpose:
To investigate the anticancer properties implicated in a natural triterpenoid (pristimerin)-induced apoptosis and inhibited proliferation in human hepatocellular carcinoma (HCC) HepG2 cell line.
Methods:
The cytotoxic activity of pristimerin in HepG2 cells was determined by MTT assay. Apoptotic morphology was observed by fluorescence microscope with Hoechst 33258 staining and percent apoptosis was measured by annexin V/PI double staining. DiOC6 for mitochondrial potential (ΔΨm) and DCFH-DA for reactive oxygen species (ROS) were determined by flow cytometry. Changes of apoptotic- related proteins were analysed by Western blot.
Results:
Pristimerin exerted a potent cytotoxic effect on HepG2 cells. After HepG2 cells were treated with pristimerin, typical apoptotic bodies, increasing the proportion of apoptotic annexin V-positive cells and activation of caspase-3 were detected in a dose-dependent manner. It was intriguing that pristimerin increased the generation of ROS with a collapse of the mitochondrial membrane potential in the cells. In addition, there was significant change in other mitochondrial membrane proteins triggered by pristimerin, such as Bcl-2 and Bax. Pristimerin also effectively induced subsequent release of cytochrome C from mitochondria into the cytosol, downregulated EGFR protein expression and inhibited downstream signaling pathways in HepG2 cells. Pretreatment with N-acetylcysteine (NAC) blocked ROS generation and resulted in loss of mitochondrial membrane potential, release of cytochrome C and apoptosis induced by pristimerin.
Conclusion:
These data indicate that ROS play an essential role in the induction of apoptosis by pristimerin in HepG2 cells.
Insights
Pristimerin, a natural compound, effectively kills liver cancer cells (HepG2) by triggering programmed cell death (apoptosis). This process involves reactive oxygen species (ROS) and mitochondrial dysfunction.
Area of Science:
- Pharmacology
- Cell Biology
- Biochemistry
Background:
- Hepatocellular carcinoma (HCC) is a major global health concern.
- Natural compounds offer potential therapeutic strategies for cancer treatment.
- Pristimerin is a triterpenoid with known biological activities.
Purpose of the Study:
- To investigate the anticancer effects of pristimerin on human hepatocellular carcinoma (HCC) HepG2 cells.
- To elucidate the mechanisms underlying pristimerin-induced apoptosis and proliferation inhibition.
Main Methods:
- Cytotoxicity assessed by MTT assay.
- Apoptosis evaluated using Hoechst staining, Annexin V/PI, and caspase-3 activation.
- Mitochondrial membrane potential (ΔΨm) and reactive oxygen species (ROS) measured by flow cytometry.
- Protein expression analyzed via Western blot.
Main Results:
- Pristimerin demonstrated potent dose-dependent cytotoxicity against HepG2 cells.
- Apoptotic morphology, increased Annexin V-positive cells, and caspase-3 activation were observed.
- Pristimerin induced ROS generation, mitochondrial membrane potential collapse, and cytochrome C release.
- Changes in Bcl-2, Bax, and EGFR expression were noted, with downstream pathway inhibition.
Conclusions:
- Pristimerin effectively induces apoptosis in HepG2 cells.
- Reactive oxygen species (ROS) play a critical role in pristimerin-mediated apoptosis.
- Mitochondrial pathways are significantly involved in pristimerin's anticancer effects.
