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Updated: Feb 1, 2026
Comparison of Three Different Methods for Determining Cell Proliferation in Breast Cancer Cell Lines
Published on: September 3, 2016
RBMS2 inhibits the proliferation by stabilizing P21 mRNA in breast cancer
1Jiangsu Breast Disease Center, the First Affiliated Hospital with Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China.
Background:
RNA binding proteins (RBPs) play an important role in regulating the metabolism of target RNAs. Aberrant expression of RBPs plays a vital role in the initiation and development of many cancers. The RBM family, which has the conserved RNA binding motif RNP1 and RNP2, shares the similar function in RNA processing and RBMS2 is a member of them. P21, also named CDKN1A, promotes cell cycle arrest and plays an important role in halting cell proliferation. In our study, we identified RBMS2 as a tumor suppressor in breast cancer. It inhibited the proliferation of breast cancer by positively regulating the stability of P21 mRNA in posttranscriptional way.
Methods:
TCGA was used to identify differentially expressed RBPs in breast cancer. The effect of RBMS2 on breast cancer proliferation was evaluated in vitro using CCK-8 assays, colony formation assays and cell-cycle assays and the in vivo effect was investigated using a mouse tumorigenicity model. The main pathway and genes regulated by RBMS2 was detected by RNA sequencing. The RNA immunoprecipitation combined with dual-luciferase reporter assay were conducted to testify the direct binding between RBMS2 and P21. Rescue assay was used to detect P21 as the main target of RBMS2.
Results:
The expression of RBMS2 was lower in breast cancer compared with normal tissues and was a favorable biomarker in breast cancer. RBMS2 inhibited the proliferation of breast cancer and P21 was the main target of RBMS2. RBMS2 stabilized the mRNA of P21 by directly binding to the AU-rich element of 3'-UTR region. Anti-proliferation activity induced by overexpression of RBMS2 was rescued by interfering with the expression of P21.
Conclusion:
In conclusion, RBMS2 acted as a tumor suppressor in breast cancer and positively regulated the expression of P21 by stabilizing its mRNA.
Insights
RBMS2 acts as a tumor suppressor in breast cancer by stabilizing P21 mRNA, inhibiting cancer cell proliferation. Lower RBMS2 expression correlates with breast cancer, making it a potential biomarker.
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- RNA binding proteins (RBPs) regulate RNA metabolism; aberrant RBP expression is linked to cancer.
- RBMS2, a member of the RBM family, functions in RNA processing.
- P21 (CDKN1A) is crucial for cell cycle arrest and halting proliferation.
Purpose of the Study:
- To investigate the role of RBMS2 in breast cancer.
- To determine if RBMS2 acts as a tumor suppressor.
- To elucidate the mechanism by which RBMS2 affects breast cancer proliferation.
Main Methods:
- TCGA data analysis to identify differentially expressed RBPs in breast cancer.
- In vitro (CCK-8, colony formation, cell-cycle assays) and in vivo (mouse tumorigenicity model) experiments to assess RBMS2's effect on proliferation.
- RNA sequencing to identify RBMS2-regulated pathways.
- RNA immunoprecipitation and dual-luciferase reporter assays to confirm RBMS2-P21 binding.
- Rescue assays to validate P21 as RBMS2's target.
Main Results:
- RBMS2 expression is significantly lower in breast cancer tissues compared to normal tissues.
- RBMS2 inhibits breast cancer cell proliferation both in vitro and in vivo.
- RBMS2 directly binds to the 3'-UTR of P21 mRNA, stabilizing it.
- Overexpression of RBMS2 inhibits proliferation, and this effect is reversed by P21 knockdown.
Conclusions:
- RBMS2 functions as a tumor suppressor in breast cancer.
- RBMS2 enhances P21 mRNA stability, thereby inhibiting breast cancer proliferation.
- RBMS2 is a potential prognostic biomarker for breast cancer.
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