Abstract
Lung adenocarcinoma (LUAD) is a subtype of lung cancer with high incidence and mortality globally. Emerging evidence suggests that circular RNAs (circRNAs) exert critical functions in human cancers, including LUAD. CircRNA_100549 (circ_100549) has been reported to be significantly upregulated in non-small cell lung cancer (NSCLC) samples, while its role in modulating LUAD progression remains to be explored. The current study aims at investigating the functional roles of circ_100549 in LUAD and its downstream molecular mechanism. First, we found that the expression of circ_100549 was higher in LUAD cell lines. Loss-of-function assays verified that depletion of circ_100549 repressed LUAD cell proliferation but accelerated cell apoptosis. Furthermore, in vivo experiments demonstrated that silencing of circ_100549 suppressed tumor growth. Subsequently, based on database analysis, we carried out a series of experiments to explore the mechanisms and effects of circ_100549 underlying LUAD progression, including RNA-binding protein immunoprecipitation (RIP), RNA/DNA pull-down, luciferase reporter, and chromatin immunoprecipitation (ChIP) assays. The results indicated that circ_100549 serves as a ceRNA by sponging miR-95-5p to upregulate BPTF expression, thus upregulating BIRC6 expression at a transcriptional level in LUAD. In summary, our study demonstrated that circ_100549 facilitates LUAD progression by upregulating BIRC6 expression.
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Data availability
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Abbreviations
- ACS:
-
American Cancer Society
- CCK-8:
-
Cell counting kit-8
- ChIP:
-
Chromatin immunoprecipitation
- circRNAs:
-
Circular RNAs
- IHC:
-
Immunohistochemistry
- IAP:
-
Inhibitors of apoptosis protein
- LUAD:
-
Lung adenocarcinoma
- MUT:
-
Mutant
- NSCLC:
-
Non-small cell lung cancer
- qPCR:
-
Quantitative polymerase chain reaction
- RIP:
-
RNA-binding protein immunoprecipitation
- siRNAs:
-
Silencing RNAs
- SCLC:
-
Small cell lung cancer
- WT:
-
Wild-type
- EdU:
-
5-Ethynyl-2′-deoxyuridine
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F.C. and J.C. co-designed this whole research. F.C., J.C., Y.Y., S.F., and J.X. performed the experiments and analyzed the data. X.X., Z.Y., and J.J. prepared the figures and wrote the manuscript. All authors have read and approved the final manuscript.
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418_2024_2275_MOESM1_ESM.tif
Transfection efficiency of circ_100549 silencing determined by qPCR. A,B The knockdown efficiency of si-circ_100549 plasmids and sh-circ_100549 plasmids was examined by qPCR. respectively. **P < 0.01. Supplementary file1 (TIF 191 KB)
418_2024_2275_MOESM2_ESM.tif
miR-95-5p inhibits tumor growth in LUAD in vitro and in vivo. A-B CCK-8 and EdU assays were carried out to assess the proliferation abilities of LUAD cells after miR-95-5p overexpressing. Scale bar: 100µm. C The apoptosis of LUAD cells transfected by miR-95-5p mimics was detected by flow cytometry analysis. D The expression of apoptosis-associated proteins Bcl-2 and Bax was measured in miR-95-5p overexpressed LUAD cells. E The volume of xenografted tumors in the mice inoculated with the indicated LUAD cells by overexpressing miR-95-5p was detected. F The weight of xenografted tumors resected from the sacrificed mice inoculated with the indicated LUAD cells was measured. G Ki67-positive expression in tumor tissues removed from the xenografts inoculated with the indicated LUAD cells was analyzed by IHC. Scale bar: 100µm. **P < 0.01. Supplementary file2 (TIF 6251 KB)
418_2024_2275_MOESM3_ESM.tif
Transfection efficiency of BPTF silencing or overexpressing and BIRC6 overexpressing determined by qPCR. A,B The silencing efficiency of si-BPTF plasmids was examined by qPCR and western blot respectively. C The efficiency of BPTF-overexpressed plasmids was detected by qPCR. D The efficiency of BIRC6-overexpressed plasmids was examined by qPCR. **P < 0.01. Supplementary file3 (TIF 452 KB)
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Chen, F., Chen, J., Yuan, Y. et al. Circ_100549 promotes tumor progression in lung adenocarcinoma through upregulation of BIRC6. Histochem Cell Biol (2024). https://doi.org/10.1007/s00418-024-02275-z
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DOI: https://doi.org/10.1007/s00418-024-02275-z