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Knockdown of AMIGO2 suppresses proliferation and migration through regulating PPAR-γ in bladder cancer
Dali Han1, Bin Xiong2, Xiangxiang Zhang3
1Department of Urology, Lanzhou University Second Hospital, Key Laboratory of Gansu Province for Urological Diseases, Clinical Center of Gansu Province for Nephro-Urology, Lanzhou University, Lanzhou, Gansu Province, China.
Purpose:
This study aims to reveal the relationship between AMIGO2 and proliferation, migration and tumorigenicity of bladder cancer, and explore the potential molecular mechanisms.
Methods:
The expression level of AMIGO2 is measured by qRT-PCR and immunohistochemistry (IHC). Stable AMIGO2 knockdown cell lines T24 and 5637 were established by lentivirus transfection. Cell Counting Kit (CCK-8 assay) was produced to determine cell proliferation, flow cytometry analysis was utilized to detect cell cycle, and wound healing assay was proceeded to test migration ability of bladder cancer cells. Xenograft mouse model was established for investigating the effect of AMIGO2 on tumor formation in vivo. The RNA Sequencing technology was applied to explore the underlying mechanisms. The expression level of PPAR-γ was measured by Western Blot.
Results:
AMIGO2 was upregulated in bladder cancer cells and tissues. Inhibited expression of AMIGO2 suppresses cell proliferation and migration. Low AMIGO2 expression inhibited tumorigenicity of 5637 in nude mice. According to RNA-Seq and bioinformatics analysis, 917 DEGs were identified. The DEGs were mainly enriched in cell-cell adhesion, peroxisome proliferators-activated receptors (PPARs) signaling pathway and some other pathways. PPAR-γ is highly expressed in bladder cancer cell lines T24 and 5637, but when AMIGO2 is knocked down in T24 and 5637, the expression level of PPAR-γ is also decreased, and overexpression of PPAR-γ could reverse the suppression effect of cell proliferation and migration caused by the inhibition of AMIGO2.
Conclusion:
AMIGO2 is overexpressed in bladder cancer cells and tissues. Knockdown of AMIGO2 suppresses bladder cancer cell proliferation and migration. These processes might be regulated by PPAR-γ signaling pathway.
Insights
AMIGO2 is overexpressed in bladder cancer, driving proliferation and migration. Inhibiting AMIGO2, possibly via the PPAR-γ pathway, suppresses tumor growth.
Area of Science:
- Oncology
- Molecular Biology
Background:
- Amphoterin-induced gene and object-2 (AMIGO2) is implicated in various cancers.
- Understanding AMIGO2's role in bladder cancer is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the relationship between AMIGO2 and bladder cancer progression (proliferation, migration, tumorigenicity).
- To explore the underlying molecular mechanisms, including the involvement of the PPAR-γ signaling pathway.
Main Methods:
- AMIGO2 expression was quantified using qRT-PCR and immunohistochemistry.
- Stable AMIGO2 knockdown cell lines were created for in vitro assays (proliferation, cell cycle, migration).
- In vivo tumorigenicity was assessed using a xenograft mouse model, and RNA sequencing identified differentially expressed genes (DEGs).
Main Results:
- AMIGO2 expression was significantly upregulated in bladder cancer tissues and cell lines.
- AMIGO2 knockdown suppressed bladder cancer cell proliferation, migration, and in vivo tumor formation.
- RNA-seq analysis revealed enrichment in cell-cell adhesion and peroxisome proliferators-activated receptors (PPARs) signaling pathways.
- PPAR-γ expression correlated with AMIGO2 levels, and its modulation affected proliferation and migration.
Conclusions:
- AMIGO2 is a key oncogene in bladder cancer, promoting proliferation and migration.
- The PPAR-γ signaling pathway is a potential mediator of AMIGO2's effects in bladder cancer.
- Targeting AMIGO2 and/or PPAR-γ may offer therapeutic strategies for bladder cancer.
