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. 2023 Oct 18;23(1):995.
doi: 10.1186/s12885-023-11515-9.

BUB1, BUB1B, CCNA2, and CDCA8, along with miR-524-5p, as clinically relevant biomarkers for the diagnosis and treatment of endometrial carcinoma

Qirong Hao  1 , Hongqin Wu  2 , Erniao Liu  2 , Lina Wang  2
Affiliations

BUB1, BUB1B, CCNA2, and CDCA8, along with miR-524-5p, as clinically relevant biomarkers for the diagnosis and treatment of endometrial carcinoma

Qirong Hao et al. BMC Cancer. .

Abstract

Background: Endometrial carcinoma (EC) is a malignant tumor of the female reproductive tract that has been associated with increased morbidity and mortality. This study aimed to identify biomarkers and potential therapeutic targets for EC.

Methods: A publicly available transcriptome data set comprising 587 EC cases was subjected to a comprehensive bioinformatics analysis to identify candidate genes responsible for EC occurrence and development. Next, we used clinical samples and cell experiments for validation.

Results: A total of 1,617 differentially expressed genes (DEGs) were identified. Analysis of patient survival outcomes revealed that BUB1, BUB1B, CCNA2, and CDCA8 were correlated with prognosis in patients with EC. Moreover, assessment of clinical samples confirmed that BUB1, BUB1B, CCNA2 and CDCA8 were strongly expressed in EC tissues. Additionally, bioinformatics and luciferase reporter assays confirmed that miR-524-5p can target and regulate these four genes. Overexpression of miR-524-5p significantly inhibited EC Ishikawa cells viability, migration and invasion. Inhibition of miR-524-5p showed the opposite results.

Conclusions: Expression of miR-524-5p reduced the migration and invasion of Ishikawa EC cells, and decreased BUB1, BUB1B, CCNA2, and CDCA8 expression. miR-524-5p, as well as BUB1, BUB1B, CCNA2, and CDCA8, may be clinically relevant biomarkers for the diagnosis and treatment of EC.

Keywords: Endometrial carcinoma (EC); Ishikawa cells, biomarker; miR-524-5p.

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Conflict of interest statement

The authors have no conflicts of interest to declare.

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Identification of DEGs associated with endometrial cancer. (A) Volcano map for DEGs identified using DESeq. (B) Volcano map for DEGs identified using edgeR. A gradual change in color from green to red indicates the gene expression transition from downregulated to upregulated. Endometrial cancer cases: 552; non-cancer cases: 35. DEGs, differentially expressed genes
Fig. 2
Fig. 2
Differential GO analysis and KEGG pathway analysis. (A) Venn diagrams showing 1617 intersecting DEGs from DESeq and edgeR screening. (B) GO enrichment analysis of the intersecting DEGs. The y-axis labels represent the aggregated GO terms. (C) KEGG pathway analysis of the intersecting DEGs. The y-axis labels represent the clustered KEGG pathway. GeneRatio represents the ratio of the number of genes enriched in a KEGG pathway to the number of DEGs that were up- or downregulated. GO, gene ontology; KEGG, kyoto encyclopedia of genes and genomes; DEGs, differentially expressed genes
Fig. 3
Fig. 3
Interaction network of central genes and proteins. (A, B) Central gene network determined using Cytoscape’s cytoHubba. The coefficients for |r| are > 0.4 and P < 0.05
Fig. 4
Fig. 4
Survival analysis of 10 genes. The Kaplan-Meier curve shows the relationship between 10 genes and the survival of EC patients. BUB1, BUB1B, PLK1, and CDCA8 were significantly related to the survival of EC patients. Patients with high AURKB, CCNB1, CCNB2, and CCNA2 expression in conjunction with high CDC20 and CDK1 expression showed poor prognosis, but it was not statistically significant. P < 0.05 indicates a statistically significant difference. Endometrial cancer cases: 552
Fig. 5
Fig. 5
miR-524-5p targeted BUB1, BUB1B, CCNA2, and CDCA8. (A) TargetScan predicted BUB1, BUB1B, CCNA2, and CDCA8 as the downstream target genes of miR-524-5p. (B) The miR-524-5p expression was detected by RT-qPCR in human normal endometrial cells (hEEC) and human endometrial cancer Ishikawa cells. (C) In TCGA database, BUB1, BUB1B, CCNA2, and CDCA8 were all increased in endometrial cancer tissues, compared with normal tissues. (D) Dual luciferase assay was used to detect the targeting of miR-524-5p with BUB1, BUB1B, CCNA2, and CDCA8 in Ishikawa cells. **P < 0.01
Fig. 6
Fig. 6
Analysis of central gene expression in 79 samples and TCGA database. (A, B) Compared with the normal group, endometrial cancer tissue showed high BUB1, BUB1B, CCNA2, and CDCA8 protein expression. P < 0.05 indicates a statistically significant difference. Endometrial cancer cases: 50; normal uterus tissue: 29. (C) Effects of miR-524-5p mimics and inhibitors in the viability of ISK cells. ISK, Ishikawa
Fig. 7
Fig. 7
Effect of miR-524-5p on wound healing, migration, and invasion of ISK cells. (A) Transfection with miR-524-5p mimic resulted in the overexpression of miR-524-5p in ISK cells. miR-524-5p overexpression reduced the (B) wound healing capacity, (C) migration ability, and (D) invasion ability of ISK cells. P < 0.05 indicates a statistically significant difference. Data are expressed as the mean ± SD of three independent experiments. The number of cells: 4 × 105/well. ISK, Ishikawa; SD, standard deviation
Fig. 8
Fig. 8
Effect of miR-524-5p on the wound healing, migration, and invasion of ISK cells. (A) Transfection with miR-524-5p inhibitors resulted in the lower expression of miR-524-5p in ISK cells. miR-524-5p inhibitors increased the (B) wound healing capacity, (C) migration ability, and (D) invasion ability of ISK cells. P < 0.05 indicates a statistically significant difference. Data are expressed as the mean ± SD of three independent experiments. Magnification of microscope: 200 ×. The number of cells: 4 × 105/well. SD, standard deviation
Fig. 9
Fig. 9
Effect of miR-524-5p on BUB1, BUB1B, CCNA2, and CDCA8 protein expression. (A–E) miR-524-5p reduced BUB1, BUB1B, CCNA2, and CDCA8 expression. Compared with the control group, the overexpression group showed reduced BUB1, BUB1B, CCNA2, and CDCA8 expression. P < 0.05 was considered statistically significant. Data are expressed as the mean ± SD of three independent experiments. The number of cells: 4 × 105/well. SD, standard deviation
Fig. 10
Fig. 10
Effect of miR-524-5p on BUB1, BUB1B, CCNA2, and CDCA8 protrein expression. (A–E) miR-524-5p inhibitors increased BUB1, BUB1B, CCNA2, and CDCA8 expression. Compared with the control group, the inhibitor miR-524-5p showed increased BUB1, BUB1B, CCNA2, and CDCA8 expression. P < 0.05 was considered statistically significant. Data are expressed as the mean ± SD of three independent experiments. The number of cells: 4 × 105/well. SD, standard deviation
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References

    1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin. 2019;69(1):7–34. - PubMed
    1. Ying J, Wang Q, Xu T, Lyu J. Establishment of a nine-gene prognostic model for predicting overall survival of patients with endometrial carcinoma. Cancer Med. 2018;7(6):2601–11. - PMC - PubMed
    1. Gaber C, Meza R, Ruterbusch JJ, Cote ML. Endometrial Cancer trends by Race and Histology in the USA: projecting the number of new cases from 2015 to 2040. J Racial Ethn Health Disparities. 2016. - PMC - PubMed
    1. Chan JK, Kapp DS, Cheung MK, Shin JY, Stieglitz D, Husain A, et al. Prognostic factors and risk of extrauterine metastases in 3867 women with grade 1 endometrioid corpus Cancer. Am J Obstet Gynecol. 2008;198(2):216e1–5. - PubMed
    1. Lowery WJ, Gehrig PA, Ko E, Secord AA, Chino J, Havrilesky LJ. Surgical staging for endometrial cancer in the elderly - is there a role for lymphadenectomy? Gynecol Oncol. 2012;126(1):12–5. - PubMed

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