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Knockdown of SETD5 Inhibits Colorectal Cancer Cell Growth and Stemness by Regulating PI3K/AKT/mTOR Pathway
Xiaohua Zhou1, Wenqiang Chen2, Duanming Zhuang3
1Department of General Surgery, Nanjing Gaochun People's Hospital, Gaochun, 211300, Jiangsu, China.
Abstract:
SET domain-containing 5 (SETD5), a member of protein lysine methyltransferase family, is expressed in multiple cancers, making it potential therapeutic targets. However, the role of SETD5 in colorectal cancer remains largely unknown. The expression of SETD5 in the 30 pairs colorectal cancer tissues samples and cell lines were determined by qRT-PCR. The functions of SETD5 was detected by knocked-down or overexpression in colorectal cancer cell lines SW480 and HCT116 cells. Cell proliferative activity, cell death, and stemness characteristics were assessed. BEZ235, a PI3K/AKT/mTOR pathway inhibitor, was used to perform rescue experiment to analyze whether SETD5 exerted its effects through activating PI3K/AKT/mTOR pathway. SETD5 was substantially upregulated in colorectal cancer, and correlated to metastasis and clinical stage of patients. Knockdown of SETD5 inhibited SW480 and HCT116 cell growth, as evidenced by the inhibition of cell viability and clone-forming. Moreover, Knockdown of SETD5 suppressed the capability of tumor sphere formation of SW480 and HCT116 cells, and reduced the expression of stemness-related proteins Nanog and Sox2. Further western blot analysis revealed that SETD5 knockdown inhibited the phosphorylation of proteins associated with the PI3K/AKT/mTOR pathway. In contrast, overexpression of SETD5 exerted the opposite effects. Mechanistically, by blocking PI3K/AKT/mTOR pathway with BEZ235, the effects of SETD5 overexpression on cell viability and Nanog and Sox2 protein expression were reversed. Our results substantiated that SETD5 functioned as an oncogene by promoting cell growth and stemness in colorectal cancer cells through activating the PI3K/AKT/mTOR signaling pathway.
Insights
SET domain-containing 5 (SETD5) is upregulated in colorectal cancer, promoting cell growth and stemness. Inhibiting SETD5 may offer a new therapeutic strategy for colorectal cancer by targeting the PI3K/AKT/mTOR pathway.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- SET domain-containing 5 (SETD5) is a protein lysine methyltransferase expressed in various cancers, but its role in colorectal cancer is unclear.
- SETD5's potential as a therapeutic target in oncology warrants investigation due to its broad expression.
Purpose of the Study:
- To investigate the role of SETD5 in colorectal cancer progression.
- To determine the functional impact of SETD5 on colorectal cancer cell growth, stemness, and the underlying molecular mechanisms.
- To explore the therapeutic potential of targeting SETD5 in colorectal cancer.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to assess SETD5 expression in colorectal cancer tissues and cell lines.
- In vitro functional assays including knockdown and overexpression of SETD5 in SW480 and HCT116 cells to evaluate cell proliferation, death, and stemness.
- Western blot analysis to examine protein expression and phosphorylation, particularly related to the PI3K/AKT/mTOR pathway.
- Rescue experiments using BEZ235, a PI3K/AKT/mTOR inhibitor, to elucidate the pathway's involvement.
Main Results:
- SETD5 expression was significantly upregulated in colorectal cancer tissues and correlated with metastasis and clinical stage.
- SETD5 knockdown inhibited colorectal cancer cell growth, viability, and sphere formation, while reducing stemness markers Nanog and Sox2.
- SETD5 knockdown suppressed the PI3K/AKT/mTOR pathway, indicated by decreased protein phosphorylation.
- SETD5 overexpression promoted cell growth and stemness, and these effects were reversed by PI3K/AKT/mTOR inhibition.
Conclusions:
- SETD5 acts as an oncogene in colorectal cancer by promoting cell proliferation and stemness.
- SETD5 activates the PI3K/AKT/mTOR signaling pathway, contributing to colorectal cancer progression.
- Targeting SETD5 or the PI3K/AKT/mTOR pathway presents a potential therapeutic strategy for colorectal cancer.
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