Abstract
Non-small cell lung cancer (NSCLC) is a common cancer with an unfavorable 5-year survival rate. We intended to explore the role of circular RNA_0074027 (circ_0074027) in NSCLC progression. The levels of circ_0074027, messenger RNA (mRNA) of its linear form paired like homeodomain 1 (PITX1), microRNA-2467-3p (miR-2467-3p) and ras homolog family member A (RHOA) mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK8) assay and plate colony assay were conducted to measure the proliferation ability of NSCLC cells. Transwell assays were used to assess cell migration and invasion abilities. Flow cytometry was utilized to analyze cell apoptosis rate. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were used to test the interaction between miR-2467-3p and circ_0074027 or RHOA. Western blot assay was performed to evaluate the protein level of RHOA in NSCLC cells. Murine xenograft model was built to evaluate the role of circ_0074027 in tumor growth in vivo. Circ_0074027 expression was prominently elevated in NSCLC tissues and cell lines. Circ_0074027 knockdown or miR-2467-3p overexpression suppressed cell proliferation, migration and invasion and facilitated cell apoptosis of NSCLC cells. Circ_0074027 interacted with miR-2467-3p, and RHOA was a target of miR-2467-3p in NSCLC cells. RHOA silencing blocked the malignant potential of NSCLC cells. Circ_0074027 silencing restrained the malignant phenotypes of NSCLC cells largely through up-regulating miR-2467-3p. Circ_0074027 knockdown notably blocked xenograft tumor growth in vivo. In conclusion, circ_0074027 accelerated NSCLC progression by binding to miR-2467-3p to induce RHOA expression.
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The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.
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Ying Liu designed and supervised the study, Zhihui Duan conducted the experiments and drafted the manuscript. Shuqing Wei involved in methodology development and analyzed the data. All authors read and approved the final manuscript.
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Clinical study was approved by the Ethics Committee of Shanxi Provincial Cancer Hospital and was carried out according to the guidelines of Declaration of Helsinki. Animal experiment was permitted by the Animal Care and Use Committee of Shanxi Provincial Cancer Hospital and performed in accordance with the guidelines of the National Animal Care and Ethics Institution.
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Supplementary Fig. 1
Circ_0074027 knockdown suppresses the EMT of NSCLC cells. (A and B) Western blot assay was employed to determine the expression of three EMT-associated markers (E-cadherin, N-cadherin and Vimentin) in A549 and H1299 cells transfected with si-NC or si-circ_0074027. **P < 0.01
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Duan, Z., Wei, S. & Liu, Y. Circ_0074027 contributes to non‐small cell lung cancer progression through positively modulating RHOA via sequestering miR-2467-3p. J Bioenerg Biomembr 53, 223–233 (2021). https://doi.org/10.1007/s10863-021-09876-6
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DOI: https://doi.org/10.1007/s10863-021-09876-6