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Thomas Flaig, MD


Study Centers on Key Role of the E3 Ligase RFWD3 in Bladder Cancer

By: Vanessa A. Carter, BS
Posted: Monday, February 6, 2023

According to Peng Jiang, MD, of the First Affiliated Hospital, Zhejiang University School of Medicine, China, and colleagues, RING finger with WD repeat domain 3 (RFWD3)—an E3 liagase—has been shown to be elevated in certain malignant tumors, but its role in the regulation of bladder cancer is unknown. A recent study, published in Histology and Histopathology, evaluated the possible role of RFWD3 in the development and progression of bladder cancer.

“In summary, RFWD3 was identified as a key regulator in the development and progression of bladder cancer, which affected cell phenotypes in vitro,” the study authors stated. “Therefore, RFWD3 may be considered as a potential therapeutic target in the treatment of bladder cancer.”

To evaluate RFWD3 expression in clinical samples, a tissue microarray containing 53 tumor tissues and 16 paracarcinoma normal tissues was used to determine its correlation with patient prognosis and tumor characteristics. Gain-of-function and loss-of-function assays were conducted using RFWD3 overexpression and knockdown cell models. Quantitative polymerase chain reaction and Western blotting were performed, as well as mean transit time, colony formation, flow cytometry, wound-healing assays, and transwell assays.

In tumor tissues, RFWD3 expression appeared to be significantly upregulated when compared with normal tissues, and patients with higher RFWD3 expression displayed a poorer prognosis and higher N stage. Moreover, bladder cancer cells with low RFWD3 expression demonstrated significantly lower proliferative activity. Wound-healing assay results further confirmed that RFWD3 knockdown decreased the mobility of bladder cancer cells.

In comparison, bladder cancer cell lines T24 and EJ experienced enhanced cell proliferation when RFWD3 was overexpressed. In addition, RFWD3 overexpression decreased the number of apoptotic cells, and it increased cell migration and colony formation of bladder cancer cells.

“This work is only the beginning of our study on the function of RFWD3 in bladder cancer, and our subsequent study will deeply explore its intrinsic mechanism and elucidate the mechanism of action,” concluded the investigators.

Disclosure: The study authors reported no conflicts of interest.

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