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Published on: May 1, 2015
[Intervention of ERRα Expression on Apoptosis Induction of Multiple Myeloma MM.1S Cells Cultured in Vitro]
Rui-Xi Zhang1, Yu-Qing Gao1, Lei Lei1
1Institute of Blood and Marrow Transplantation, Medical College of Soochow University; The First Affiliated Hospital of Soochow University; Jiangsu Institute of Hematology;Collaborative Innovation Center of Hematology ;Suzhou 215000, Jiangsu Province, China.
Objective:
To investigate the effect of two different approaches ERRα strategy on the apoptosis in multiple myeloma cell line MM.1S.
Methods:
For the one strategy, shRNA was mediated by lentivirus. Stable cell clones were established by transfecting the lentivirus into MM.1S cells and screened by puromycin. For the other strategy, XCT790, a specific reverse agonist of ERRα, was used to treat MM.1S cells. The apoptosis of the cells was analyzed by flow cytometry after ERRα was down-regulated. Western blot assay was used to detect the apoptosis of related proteins.
Results:
The knocked down ERRα was achieved, lentivirus with shERRα were successfully infected into MM.1S and ERRα was reduced significantly. Knockdown of ERRα could induce MM.1S cell apoptosis dramatically. Meanwhile, the expression of cleaved PARP (a kind of apoptosis related markers) was significantly increased following depletion of ERRα in MM.1S cells. XCT790 could significantly down-regulate the expression of ERRα protein in MM.1S cells, which was consistent with the effect caused by shRNA.
Conclusion:
Interference the expression of ERRα by shRNA or XCT790 can induce apparent apoptosis in MM.1S cells, which indicating that ERRα is crucial for the survival of myeloma cells.
Insights
Estrogen-related receptor alpha (ERRα) is crucial for myeloma cell survival. Down-regulating ERRα expression through shRNA or XCT790 significantly induces apoptosis in multiple myeloma cells.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Multiple myeloma is a hematologic malignancy.
- Estrogen-related receptor alpha (ERRα) plays a role in cancer cell survival.
- Targeting ERRα presents a potential therapeutic strategy.
Purpose of the Study:
- To investigate the impact of ERRα down-regulation on apoptosis in the MM.1S multiple myeloma cell line.
- To compare two distinct methods for reducing ERRα expression: shRNA and the small molecule XCT790.
Main Methods:
- Lentiviral-mediated shRNA delivery to stably down-regulate ERRα in MM.1S cells.
- Treatment of MM.1S cells with XCT790, a specific reverse agonist of ERRα.
- Apoptosis analysis using flow cytometry and Western blot to detect apoptosis-related proteins, including cleaved PARP.
Main Results:
- Successful significant reduction of ERRα expression was achieved using both shRNA and XCT790.
- Down-regulation of ERRα markedly induced apoptosis in MM.1S cells.
- Increased expression of cleaved PARP, a marker of apoptosis, was observed following ERRα depletion.
Conclusions:
- Interference with ERRα expression via shRNA or XCT790 effectively induces apoptosis in MM.1S cells.
- These findings highlight ERRα as a critical factor for the survival of myeloma cells.
- Targeting ERRα represents a promising therapeutic avenue for multiple myeloma treatment.
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