Long non-coding RNA ARHGAP5-AS1 inhibits migration of breast cancer cell via stabilizing SMAD7 protein
Chen-Long Wang1,2, Jing-Chi Li1, Ci-Xiang Zhou1
1Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education, Shanghai Jiao Tong University School of Medicine (SJTU-SM), No. 280, Chong Qing South Rd, Shanghai, 200025, China.
Purpose:
Tumor metastasis is the main cause of death from breast cancer patients and cell migration plays a critical role in cancer metastasis. Recent studies have shown long non-coding RNAs (lncRNAs) play an essential role in the initiation and progression of cancer. In the present study, the role of an LncRNA, Rho GTPase Activating Protein 5- Antisense 1 (ARHGAP5-AS1) in breast cancer was investigated.
Methods:
RNA sequencing was performed to find out dysregulated LncRNAs in MDA-MB-231-LM2 cells. Transwell migration assays and F-actin staining were utilized to estimate cell migration ability. RNA pulldown assays and RNA immunoprecipitation were used to prove the interaction between ARHGAP5-AS1 and SMAD7. Western blot and immunofluorescence imaging were used to examine the protein levels. Dual luciferase reporter assays were performed to evaluate the activation of TGF-β signaling.
Results:
We analyzed the RNA-seq data of MDA-MB-231 and its highly metastatic derivative MDA-MB-231-LM2 cell lines (referred to as LM2) and identified a novel lncRNA (NR_027263) named as ARHGAP5-AS1, which expression was significantly downregulated in LM2 cells. Further functional investigation showed ARHGAP5-AS1 could inhibit cell migration via suppression of stress fibers in breast cancer cell lines. Afterwards, SMAD7 was further identified to interact with ARHGAP5-AS1 by its PY motif and thus its ubiquitination and degradation was blocked due to reduced interaction with E3 ligase SMURF1 and SMURF2. Moreover, ARHGAP5-AS1 could inhibit TGF-β signaling pathway due to its inhibitory role on SMAD7.
Conclusion:
ARHGAP5-AS1 inhibits breast cancer cell migration via stabilization of SMAD7 protein and could serve as a novel biomarker and a potential target for breast cancer in the future.
Insights
Rho GTPase Activating Protein 5- Antisense 1 (ARHGAP5-AS1) inhibits breast cancer cell migration by stabilizing SMAD7 protein. This lncRNA shows potential as a biomarker and therapeutic target for breast cancer treatment.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- Tumor metastasis is a primary cause of breast cancer mortality.
- Long non-coding RNAs (lncRNAs) are increasingly recognized for their roles in cancer initiation and progression.
Purpose of the Study:
- To investigate the role of the lncRNA ARHGAP5-AS1 in breast cancer.
- To elucidate the molecular mechanisms by which ARHGAP5-AS1 influences breast cancer cell migration.
Main Methods:
- RNA sequencing to identify dysregulated lncRNAs in metastatic breast cancer cells.
- Transwell migration assays and F-actin staining to assess cell migration.
- RNA pulldown, RNA immunoprecipitation, Western blot, immunofluorescence, and dual luciferase reporter assays to determine molecular interactions and signaling pathway activation.
Main Results:
- ARHGAP5-AS1 was found to be significantly downregulated in highly metastatic breast cancer cells.
- ARHGAP5-AS1 inhibits breast cancer cell migration by suppressing stress fibers.
- ARHGAP5-AS1 stabilizes SMAD7 protein by blocking its ubiquitination and degradation, thereby inhibiting the TGF-β signaling pathway.
Conclusions:
- ARHGAP5-AS1 inhibits breast cancer cell migration through SMAD7 protein stabilization.
- ARHGAP5-AS1 presents potential as a novel biomarker and therapeutic target for breast cancer.
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