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Circ_0000274 contributes to renal cell carcinoma progression by regulating miR-338-3p/NUCB2 axis and JAK1/STAT3

Qiangyuan Qi¹, Yingying Sun², Ying Yang³

¹Department of Urology Surgery, Linyi Central Hospital, Linyi 276400, Shandong Province, China.

Transplant Immunology

|May 15, 2022

Abstract

Background:

Kidney transplant recipients (KTRs) are at increased risk of developing renal cell carcinoma (RCC). Accumulating evidence has demonstrated that circular RNAs (circRNAs) are essential players in tumor advancement. However, the functions of circ_0000274 in renal cell carcinoma (RCC) are barely explored.

Methods:

The primary RCC cell lines 786-O and A498 were used in this study. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was employed for the RNA levels of circ_0000274, microRNA-338-3p (miR-338-3p) and nucleobindin 2 (NUCB2). RNase R assay was conducted to analyze the feature of circ_0000274.Cell Counting Kit-8 (CCK-8) assay, colony formation assay, transwell assay, tube formation assay and flow cytometry analysis were conducted for cell viability, colony formation, metastasis, angiogenesis and apoptosis, respectively. Western blot assay was utilized for protein levels. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were adopted to analyze the associations of circ_0000274 RNA, miR-338-3p RNA and NUCB2 protein. Murine xenograft model was established to explore the function of circ_0000274 RNA in vivo. Immunohistochemistry (IHC) assay was used to analyze NUCB2 protein level in xenograft tumors.

Results:

Compared to normal tissues and cells, circ_0000274 RNA level was elevated in RCC tissues and cells. Knockdown of circ_0000274 RNA suppressed cell viability, colony formation, metastasis and tube formation and promoted apoptosis in RCC cells in vitro. Circ_0000274 RNA sponged miR-338-3p RNA to positively regulate NUCB2 protein in RCC cells. Inhibition of miR-338-3p reversed the impacts of circ_0000274 knockdown on RCC cell malignant behaviors. MiR-338-3p RNA overexpression repressed the malignant phenotypes of RCC cells, while NUCB2 protein elevation could abrogate the effect. Moreover, circ_0000274 RNA knockdown blocked tumorigenesis in vivo. Besides, circ_0000274 RNA knockdown inactivated the JAK1/STAT3 protein signaling pathway.

Conclusion:

Circ_0000274 RNA functioned as an oncogene in RCC development by regulating miR-338-3p RNA/NUCB2 protein axis and activating the JAK1/STAT3 protein signaling pathway.

Insights

Circular RNA circ_0000274 acts as an oncogene in renal cell carcinoma (RCC) by promoting tumor growth and metastasis. Its inhibition suppresses RCC progression via the miR-338-3p/NUCB2 pathway and JAK1/STAT3 signaling.

Area of Science:

Oncology
Molecular Biology
Genetics

Background:

Kidney transplant recipients (KTRs) face an elevated risk of renal cell carcinoma (RCC).
Circular RNAs (circRNAs) are increasingly recognized for their role in cancer progression.
The specific function of circ_0000274 in RCC remains largely uninvestigated.

Purpose of the Study:

To investigate the role and mechanism of circ_0000274 in the development of renal cell carcinoma (RCC).
To explore the potential of targeting circ_0000274 as a therapeutic strategy for RCC.

Main Methods:

Quantitative real-time polymerase chain reaction (qRT-PCR) and RNase R assay to analyze RNA levels and features.
In vitro assays (CCK-8, colony formation, transwell, tube formation, flow cytometry) to assess cell viability, metastasis, angiogenesis, and apoptosis.
In vivo studies using a murine xenograft model to evaluate tumorigenesis.
Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays to elucidate molecular interactions.

Main Results:

Circ_0000274 expression was significantly upregulated in RCC tissues and cell lines.
Knockdown of circ_0000274 inhibited RCC cell proliferation, migration, invasion, and angiogenesis while promoting apoptosis.
Circ_0000274 acts as a sponge for microRNA-338-3p (miR-338-3p), leading to increased nucleobindin 2 (NUCB2) protein expression.
Inhibition of miR-338-3p reversed the anti-tumor effects of circ_0000274 knockdown.
Overexpression of NUCB2 partially abrogated the tumor-suppressive effects of miR-338-3p.
Circ_0000274 knockdown suppressed tumor growth in vivo and inactivated the JAK1/STAT3 signaling pathway.

Conclusions:

Circ_0000274 functions as an oncogene in RCC development.
The oncogenic role of circ_0000274 is mediated through the miR-338-3p/NUCB2 axis.
Circ_0000274 activates the JAK1/STAT3 signaling pathway, contributing to RCC progression.

Read full analysis

Keywords:

JAK1/STAT3 NUCB2 RCC circ_0000274 miR-338-3p

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