[Lonidamine induces apoptosis via endoplasmic reticulum stress response and down-regulating cIAP expression in human
Furong Shao1, Liang Wang, Xiaoqin Chu
1School of Nursing, Anhui University of Chinese Medicine, Hefei 230038, China.
Objective:
To investigate the effect of lonidamine on apoptosis of human breast carcinoma cells MCF-7 and the possible mechanisms.
Methods:
MTT assay and colony-forming assay were used to evaluate the growth inhibition induced by lonidamine in breast cancer MCF-7 cells. PI/Annexin-V staining was used to detect the apoptotic cells. The ATP levels in the cells were detected using an ATP assay kit. The expression of glucose regulated protein 78 (GRP78), inhibitor of apoptosis protein (cIAP1) and caspase-8 were analyzed with Western blotting.
Results:
MTT assay and colony-forming assay showed that 50-250 mmol/L lonidamine caused a time- and concentration-dependent inhibition of MCF-7 cell proliferation. Exposure to increased concentrations of lonidamine resulted in significantly increased apoptosis rate in MCF-7 cells. In MCF-7 cells treated with 50, 150 and 250 mmol/L lonidamine for 5 h, the intracellular ATP levels were lowered to 80.67%, 62.78% and 30.73% of the control level, respectively. Western blotting showed that lonidamine up-regulated the expression of GRP78, down-regulated the expression of cIAP1 and promoted caspase-8 activation as the treatment time extended.
Conclusion:
Lonidamine can inhibit the proliferation and induce apoptosis in MCF-7 cells, and these effects are probably mediated by reducing ATP level, inducing endoplasmic reticulum stress response, down-regulating cIAP1, and promoting caspase-8 activation in the cells.
Insights
Lonidamine inhibits human breast cancer cell growth by inducing apoptosis. This effect is linked to reduced ATP levels, endoplasmic reticulum stress, and altered protein expression, including caspase-8 activation.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Human breast carcinoma MCF-7 cells are a common model for studying breast cancer.
- Understanding the mechanisms of anti-cancer drugs is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the effects of lonidamine on MCF-7 cell apoptosis.
- To elucidate the underlying molecular mechanisms of lonidamine's action.
Main Methods:
- Cell proliferation assessed by MTT and colony-forming assays.
- Apoptosis detected using PI/Annexin-V staining.
- Intracellular ATP levels measured with an ATP assay kit.
- Protein expression (GRP78, cIAP1, caspase-8) analyzed by Western blotting.
Main Results:
- Lonidamine demonstrated dose- and time-dependent inhibition of MCF-7 cell proliferation.
- Increased lonidamine concentrations significantly elevated the apoptosis rate.
- Lonidamine reduced intracellular ATP levels and modulated GRP78, cIAP1, and caspase-8 expression.
Conclusions:
- Lonidamine effectively inhibits MCF-7 cell proliferation and induces apoptosis.
- Mechanisms involve ATP depletion, endoplasmic reticulum stress, cIAP1 downregulation, and caspase-8 activation.
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