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Modeling Oral-Esophageal Squamous Cell Carcinoma in 3D Organoids
Published on: December 23, 2022
POLE2 knockdown reduce tumorigenesis in esophageal squamous cells
Yongjun Zhu1, Gang Chen1, Yang Song1
1Department of Cardiothoracic Surgery, Huashan Hospital, Fudan University, No. 12, Mid, Wulumuqi Rd, Shanghai, China.
Background:
Esophageal squamous cell carcinoma (ESCC) is one of the most frequent malignant tumors originated from digestive system around the world and the treatment was limited by the unclear mechanism. DNA polymerase epsilon 2, accessory subunit (POLE2) is involved in DNA replication, repair, and cell cycle control, whose association with ESCC is still not clear.
Methods:
In this study, the expression level of POLE2 in ESCC tissues was detected by IHC. The POLE2 knockdown cell line was constructed, identified by qPCR and western blot and used for detecting cellular functions and constructing xenotransplantation mice model. MTT Assay, colony formation assay, flow cytometry, wound-healing assay and Transwell assay were used to detected cell proliferation, apoptosis and migration.
Results:
We firstly identified that the expression of POLE2 was overexpressed in ESCC. Moreover, the high expression of POLE2 can predict the tumor deterioration and poor prognosis of ESCC patients. Additionally, downregulation of POLE2 was involved in ESCC progression by promoting proliferation, migration, and inhibiting apoptosis in vitro. In vivo studies proved that POLE2 was positively correlated with ESCC tumor formation, which was consistent with the results in vitro. We also illuminated that POLE2 knockdown upregulated pro-apoptotic proteins (Bax, Caspase3, CD40L, FasL, IGFBP-5 and P21) and downregulated anti-apoptotic proteins (CLAP-2, IGF-I and sTNF-R2). In addition, POLE2 was involved in ESCC via targeting PI3K/Akt, Cyclin D1 signaling pathway.
Conclusions:
Therefore, POLE2 was proved to be involved in the development of ESCC, which may be a potential therapeutic target and bring new breakthroughs in the treatment of ESCC.
Insights
DNA polymerase epsilon 2, accessory subunit (POLE2) is overexpressed in esophageal squamous cell carcinoma (ESCC), promoting tumor progression. Downregulating POLE2 inhibits ESCC cell proliferation and migration, suggesting POLE2 as a potential therapeutic target for ESCC.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Esophageal squamous cell carcinoma (ESCC) is a prevalent digestive system malignancy with unclear underlying mechanisms.
- DNA polymerase epsilon 2, accessory subunit (POLE2) plays roles in DNA replication, repair, and cell cycle control.
- The specific involvement of POLE2 in ESCC pathogenesis remains largely unelucidated.
Purpose of the Study:
- To investigate the expression levels of POLE2 in ESCC tissues.
- To determine the correlation between POLE2 expression and ESCC patient prognosis.
- To elucidate the functional role of POLE2 in ESCC progression both in vitro and in vivo.
Main Methods:
- Immunohistochemistry (IHC) was employed to assess POLE2 expression in ESCC tissues.
- POLE2 knockdown cell lines were established and validated using qPCR and Western blot.
- Cellular proliferation, apoptosis, migration, and tumor formation in xenotransplantation models were evaluated using MTT assays, colony formation assays, flow cytometry, wound-healing assays, and Transwell assays.
Main Results:
- POLE2 was found to be significantly overexpressed in ESCC tissues, correlating with tumor progression and poor patient prognosis.
- Downregulation of POLE2 suppressed ESCC cell proliferation and migration while promoting apoptosis in vitro.
- In vivo studies confirmed that POLE2 positively influenced ESCC tumor formation, with its knockdown altering the expression of key apoptosis-related proteins and impacting the PI3K/Akt and Cyclin D1 signaling pathways.
Conclusions:
- POLE2 is implicated in the development and progression of ESCC.
- POLE2 may serve as a valuable prognostic biomarker for ESCC.
- Targeting POLE2 presents a potential therapeutic strategy for improving ESCC treatment outcomes.
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