Apatinib Inhibits Stem Properties and Malignant Biological Behaviors of Breast Cancer Stem Cells by Blocking Wnt/β-catenin Signal Pathway through Downregulating LncRNA ROR

Background: Cancer stem cells could influence tumor recurrence and metastasis.

Objective: To develop a new effective treatment modality targeting breast cancer stem cells (BCSCs) and to explore the role of Apatinib in BCSCs.

Methods: BCSCs were isolated from MDA-MB-231 cells by the immune magnetic beads method. BCSCs were treated with Apatinib, lentiviral plasmids (lncRNA ROR), and iCRT-3 (Wnt pathway inhibitors). Viability, colony numbers, sphere numbers, apoptosis, migration, invasion of BCSCs were detected by MTT, colony formation, tumorsphere, flow cytometry, wound-healing, transwell assays, respectively. The expressions of markers (ABCG2, CD44, CD90, and CD24), epithelial-mesenchymal transition (EMT)-related molecules (Ecadherin, N-cadherin, Vimentin, MMP-2, MMP-9), and Wnt/β-catenin pathway-related proteins (Wnt3a, Wnt5a, β-catenin) in breast cancer stem cells were determined by performing Western blot and qRT-PCR analysis.

Results: Apatinib decreased the viability and colony numbers of BCSCs in a concentration-dependent manner, and it also reduced sphere numbers, suppressed migration, invasion and lncRNA ROR expression, and induced apoptosis of BCSCs. However, these results were partially reversed by lncRNA ROR overexpression. Apatinib suppressed stem property, EMT process, and Wnt/β-catenin pathway in BCSCs, which was partially reversed by lncRNA ROR overexpression. Moreover, lncRNA ROR overexpression increased the colony and sphere numbers and promoted the cell viability, apoptosis inhibition, migration, and invasion of BCSCs, but these effects were partially reversed by iCRT-3. LncRNA ROR overexpression increased the stem property, EMT process, and Wnt/β-catenin pathway, which were partially counteracted by iCRT-3.

Conclusion: Apatinib inhibited stem property and malignant biological behaviors of BCSCs by blocking the Wnt/β-catenin signal pathway through down-regulating lncRNA ROR.

 

Comments:

The study aimed to develop a new treatment approach targeting breast cancer stem cells (BCSCs) and investigate the role of Apatinib, a drug, in BCSCs. The researchers used the MDA-MB-231 cell line to isolate BCSCs using the immune magnetic beads method. They then treated the BCSCs with Apatinib, lentiviral plasmids (lncRNA ROR), and iCRT-3 (Wnt pathway inhibitors) to evaluate their effects.

The researchers conducted various assays to assess the impact of the treatments on BCSCs. They measured viability, colony formation, sphere formation, apoptosis, migration, and invasion of BCSCs using MTT, colony formation, tumorsphere, flow cytometry, wound-healing, and transwell assays, respectively. Additionally, they analyzed the expression of specific markers (ABCG2, CD44, CD90, and CD24), epithelial-mesenchymal transition (EMT)-related molecules (E-cadherin, N-cadherin, Vimentin, MMP-2, MMP-9), and Wnt/β-catenin pathway-related proteins (Wnt3a, Wnt5a, β-catenin) in BCSCs using Western blot and qRT-PCR analysis.

The results showed that Apatinib reduced the viability and colony numbers of BCSCs in a concentration-dependent manner. It also decreased sphere formation, inhibited migration and invasion, suppressed the expression of lncRNA ROR, and induced apoptosis in BCSCs. However, the effects of Apatinib were partially reversed when lncRNA ROR was overexpressed. Apatinib also suppressed the stem-like properties, the process of epithelial-mesenchymal transition (EMT), and the Wnt/β-catenin pathway in BCSCs, and these effects were partially reversed by lncRNA ROR overexpression. Moreover, lncRNA ROR overexpression increased colony and sphere formation, enhanced cell viability, inhibited apoptosis, promoted migration and invasion in BCSCs. However, these effects were partially reversed by iCRT-3, which is a Wnt pathway inhibitor. LncRNA ROR overexpression also increased stem-like properties, EMT process, and Wnt/β-catenin pathway activation, but these effects were partially counteracted by iCRT-3.

In conclusion, the study suggests that Apatinib inhibits the stem-like properties and malignant behavior of BCSCs by blocking the Wnt/β-catenin signaling pathway through downregulation of lncRNA ROR. This research provides insights into the potential of Apatinib as a treatment modality targeting BCSCs and highlights the role of the Wnt/β-catenin pathway and lncRNA ROR in BCSCs.