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Published on: March 30, 2019
circ-ATAD1 as Competing Endogenous RNA for miR-191-5p Forces Non-small Cell Lung Cancer Progression
Zhihua Wan1, Shanshan Jia2, Junhua Lu2
1Baoding First Central Hospital, Baoding City, 071000, Hebei Province, China.
Abstract:
The association of circular RNAs (circRNAs) with non-small cell lung cancer (NSCLC) has been recognized extensively. In view of this, our study particularly surveyed the underlying mechanism of circ-ATAD1 in the disease. First, an analysis of the clinical expression of circ-ATPase family AAA domain containing 1 (ATAD1) was performed, followed by further evaluation of the relationship between circ-ATAD1 expression and prognosis. Then, A549 cells were treated with single transfection or combined transfection with the plasmid vectors that interfere with circ-ATAD1 or miR-191-5p. circ-ATAD1 and miR-191-5p levels were detected by reverse transcription quantitative polymerase chain reaction to verify the transfection success. Then, cell proliferation was checked by cell count kit-8 and clonal formation test. Cell apoptosis was analyzed by flow cytometry. Cell migration and invasion were examined by wound healing assay and Transwell. Finally, the targeting of miR-191-5p to circ-ATAD1 or Forkhead Box K1 (FOXK1) was verified by bioinformation website starBase analysis and dual-luciferase reporter assay. circ-ATAD1 was expressed abundantly in tumor tissues of NSCLC patients and had a predictive value in poor prognosis. circ-ATAD1 underexpression or miR-191-5p overexpression could obstruct A549 cells to behave aggressively, while circ-ATAD1 upregulation or miR-191-5p depletion resulted in the promotion of aggressiveness of A549 cells. Interestingly, circ-ATAD1 could decoy miR-191-5p. miR-191-5p negatively regulated FOXK1 expression, and downregulating miR-191-5p or upregulating FOXK1 rescued circ-ATAD1 downregulation-mediated influences on NSCLC cells. circ-ATAD1 accelerates NSCLC progression by absorbing miR-191-5p to upregulate FOXK1 expression.
Insights
Circular RNA circ-ATAD1 promotes non-small cell lung cancer (NSCLC) progression by sponging miR-191-5p, leading to increased Forkhead Box K1 (FOXK1) expression. This mechanism highlights circ-ATAD1 as a potential therapeutic target for NSCLC.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- Circular RNAs (circRNAs) are increasingly recognized for their roles in various cancers, including non-small cell lung cancer (NSCLC).
- The specific mechanisms by which circRNAs influence NSCLC progression require further elucidation.
Purpose of the Study:
- To investigate the role and mechanism of circ-ATAD1 in non-small cell lung cancer (NSCLC).
- To explore the relationship between circ-ATAD1 expression and NSCLC patient prognosis.
- To elucidate the regulatory pathway involving circ-ATAD1, miR-191-5p, and FOXK1 in NSCLC cells.
Main Methods:
- Clinical expression analysis of circ-ATAD1 in NSCLC tissues and correlation with prognosis.
- In vitro experiments using A549 cells with circ-ATAD1 and miR-191-5p manipulation.
- Assays for cell proliferation (CCK-8, colony formation), apoptosis (flow cytometry), migration (wound healing), and invasion (Transwell); verification of molecular interactions using dual-luciferase reporter assay and starBase analysis.
Main Results:
- Circ-ATAD1 was significantly upregulated in NSCLC tissues and associated with poor prognosis.
- Circ-ATAD1 overexpression promoted NSCLC cell proliferation, migration, and invasion, while its downregulation inhibited these processes.
- Circ-ATAD1 acts as a sponge for miR-191-5p, which negatively regulates FOXK1. Upregulation of FOXK1 or downregulation of miR-191-5p rescued the effects of circ-ATAD1 downregulation.
Conclusions:
- Circ-ATAD1 accelerates NSCLC progression by sequestering miR-191-5p, thereby upregulating Forkhead Box K1 (FOXK1) expression.
- Circ-ATAD1 serves as a potential biomarker for NSCLC prognosis and a therapeutic target.
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