Abstract
Purpose
F-box proteins, as components of the Skp1-Cullin 1-F-box protein (SCF) E3 ubiquitin ligase, can specifically bind to substrates and regulate multiple tumor behaviors. However, the role of F-box proteins in squamous-cell lung carcinoma (SqCLC) has not been established.
Methods
We identified the differentially expressed F-box protein-encoding genes in SqCLC by analyzing data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Prognosis data were evaluated using the Kaplan–Meier (KM) plotter website. The FBXO5 and FBXO45 mRNA levels were analyzed by real time RT-PCR. The impact of the inhibition of these genes with si-RNA on apoptosis and migration was also investigated.
Results
The FBXO45 and FBXO5 genes were significantly up-regulated in SqCLC compared with normal lung (p values = 0.002 and 0.025, respectively). FBXO45 was significantly elevated in each tumorigenic step, including dysplasia, in situ and SqCLC. The RT-PCR analysis results showed that FBXO5 and FBXO45 were elevated in cancer tissues (p values = 0.024 and 0.004, respectively). Overexpression of FBXO5 and FBXO45 was associated with shorter overall survival (OS) in the SqCLC patients from the K–M plotter database [FBXO5 HR: 1.53 (1.03–2.28), p = 0.036]; [FBXO45 HR: 1.47 (1.03–2.08), p = 0.030]. The GO and KEGG pathway analysis showed that FBXO5 and FBXO45 were associated with cell cycle and adhesion, respectively. Knockdown of FBXO5 leads to increased apoptosis, while knockdown of FBXO45 facilitates the process of epithelial–mesenchymal transition (EMT).
Conclusions
Our results provide evidence that FBXO45 and FBXO5 may play a key role in tumorigenesis and prognosis of SqCLC.
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Funding
This study was funded by National Natural Science Foundation of China (8160101782), and Key Research and Development Plan of Shandong Province (2015GSF118063).
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Jiajun Du declares that he has no conflict of interest. Kai Wang declares that he has no conflict of interest. Xiao Qu declares that he has no conflict of interest. Shaorui Liu declares that he has no conflict of interest. Xudong Yang declares that he has no conflict of interest. Fenglong Bie declares that he has no conflict of interest. Yu Wang declares that she has no conflict of interest. Cuicui Huang declares that she has no conflict of interest.
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All procedures performed in studies involving human participants were in accordance with the ethical standards of the ethic community of Shandong Provincial Hospital and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
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432_2018_2653_MOESM1_ESM.tif
Mutation alterations of F-box proteins in all TCGA LUSC samples from cBioportal (S1A). Detailed mutations of FBXW7 were shown in S1B. Kaplan Meier curve from K–M plotter showed that over-expression of FBXW7 indicated a favorable prognosis (S1C) (TIF 2332 KB)
432_2018_2653_MOESM2_ESM.tif
The impact of differentially expressed F-box proteins on overall survival in I-IIIA resected SqCLC patients from TCGA database (FBXL6: A; FBXL19: B; FBXO5: C; FBXO17: D; FBXO27: E; FBXO32: F; FBXO41: G; FBXO43: H; FBXO45: I) (TIF 596 KB)
432_2018_2653_MOESM3_ESM.tif
Protein-protein interaction (PPI) network of FBXO45 (S3A) and FBXO5 (S3B) in TCGA SqCLC database. GSEA analysis showed pathways enriched in high FBXO5 group (S3C), low FBXO45 group (S3D) and high FBXO45 group (S3E) (TIF 1194 KB)
432_2018_2653_MOESM4_ESM.tif
The expression of FBXO5 and FBXO45 was elevated in squamous lung cancer cell lines than lung adenocarcinoma cell lines (GSE57083, S4A). Protein expression of FBXO5 and FBXO45 in cell lines was shown in S4B. Immunohistochemical (IHC) images showed FBXO5 staining of pneumonocytes in normal lung tissues (Upper line) and FBXO5 staining in squamous lung cancer tissues (Bottom line, from left to right: not detected, weak staining, moderate/strong staining, S4E). Immunohistochemical (IHC) images showed FBXO45 staining of pneumonocytes in normal lung tissues (Upper line) and FBXO45 staining in squamous lung cancer tissues (Bottom line, from left to right: not detected, weak staining, moderate/strong staining, S4F). The quantitive scores were shown in S4C. Wound healing assay results showed the increased migration of FBXO5 knockdown group (bottom line) compared to negative group (upper line) at 0, 24, 48 hours (from left to right, S4D) (TIF 14462 KB)
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Wang, K., Qu, X., Liu, S. et al. Identification of aberrantly expressed F-box proteins in squamous-cell lung carcinoma. J Cancer Res Clin Oncol 144, 1509–1521 (2018). https://doi.org/10.1007/s00432-018-2653-1
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DOI: https://doi.org/10.1007/s00432-018-2653-1