Diagnostic and prognostic value of RUNX1 in ovarian serous cystadenocarcinoma and its role in cancer progression via suppressing ferroptosis

Deng He; Yu Zhou; Tiantian Feng; Hu Wang; Nenghuan Tang; Shangqi Ni; Wang Xi; Chengju Zhang

doi:10.2478/ahem-2026-0003

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Diagnostic and prognostic value of RUNX1 in ovarian serous cystadenocarcinoma and its role in cancer progression via suppressing ferroptosis

Postępy Higieny i Medycyny Doświadczalnej

Volume 80 (2026): Issue 1 (January 2026)

By: Deng He , Yu Zhou , Tiantian Feng , Hu Wang , Nenghuan Tang , Shangqi Ni , Wang Xi and Chengju Zhang

Open Access

|Mar 2026

Figures & Tables

RUNX1 is an independent prognostic factor associated with both elevated expression in tumor tissues and poor clinical outcomes in OSC patients. (a) The expression levels of the top ten prognosis-related genes in OSC tissues (n = 427) and normal controls (n = 88). Data processing method: log2 (value + 1). Data were analyzed by unpaired two-tailed Student’s t-test. Data processing method: log₂(value + 1). ***p < 0.001. (b) Forest plot of univariate Cox regression analysis for OS. Univariate Cox proportional hazards regression was performed using the survival R package (v3.3.1). p < 0.05 was considered statistically significant. (c) Forest plot of multivariate Cox regression analysis for OS, HR, and 95% CI are shown. Statistical significance was assessed by the Wald test

Diagnostic and prognostic significance of RUNX1 in patients with OSC. (a) ROC analysis of RUNX1 in OSC patients. The AUC for RUNX1 was 0.706 (DeLong’s test, p < 0.001). (b) Data were extracted in TPM format alongside corresponding clinical records. Kaplan–Meier survival curves comparing OS between OSC patients with high (n = 190) vs low (n = 189) RUNX1 expression. (c) Prognostic nomogram integrating RUNX1 expression and clinical variables to predict 1-, 3-, and 5-year OS probability

The analysis of top ten hub genes and the expression of the Top ten hub genes in OSC. (a) PPI analysis of the network linking RUNX1 in homo sapiens via STRING database (confidence threshold >0.400). (b) The Top ten hub gene linking RUNX1 in homo sapiens was shown by Cytoscape (CytoHubba plugin, ranked by MCC method)

RUNX1 is potentially involved in regulating ferroptosis in OSC cells. (a) Venn diagram. Differential analysis was performed using DESeq2 package [42,43] on raw counts matrices from public datasets. Protein-coding genes meeting |log2FoldChange| >0.5 and adjusted p < 0.05 were filtered. Intersection was obtained with ferroptosis-related gene sets from GeneCards database. (b) GO/KEGG enrichment analysis combined with FC criteria for the 194 intersecting genes. Visualization was implemented using ggplot2 package to generate Chord diagram. (c) Correlation analysis of ferroptosis-related genes and RUNX1. RNA-seq data in TPM format processed via STAR workflow and clinical data from TCGA-OV project (n = 381). Data processing method: log2 (value + 1). Spearman analysis was performed

Knockdown of RUNX1 inhibits cell proliferation in SHIN-3 and OVCAR-3. (a) SHIN-3 and OVCAR-3 cells were infected with lentiviruses carrying RUNX1 shRNA or NC shRNA and assessed at 24, 48, and 72 h post-infection. Cell viability was measured by MTT assay following RUNX1 knockdown. Data are presented as mean value ± SD of at least three independent biological replicates, each with three replicates. Two-way analysis of variance (ANOVA) followed by Tukey s multiple comparisons test was used to compare the RUNX1 shRNA group with the NC shRNA group at each time point. **p < 0.01 vs NC shRNA group at each time point. (b) EdU incorporation assay with immunofluorescence staining. SHIN-3 cells infected with RUNX1 shRNA or NC shRNA lentiviruses were photographed to visualize EdU-positive cells at 48 h. Data are mean value ± SD from five randomly selected fields per well across three independent experiments. **p < 0.01. (c) Colony formation assay in RUNX1-knockdown SHIN-3 cells. Cells infected with RUNX1 shRNA or control shRNA lentiviruses were cultured for 2 weeks, with colonies (≥ 50 cells) counted and displayed. Data are mean value ± SD from three independent experiments, each performed in triplicates. **p < 0.01

Knockdown of RUNX1 enhances erastin-induced ferroptosis in SHIN-3 and OVCAR-3 cells. (a) MTT assay. RUNX1 expression was knocked down by infection with RUNX1-specific shRNA lentiviral particles and treated with increasing erastin concentrations for 48 h. Viability was determined in RUNX1 shRNA or Ctrl.shRNA lentivirus-infected SHIN-3 and OVCAR-3 cells using MTT assay. Data are mean value ± SD of three independent experiments. **p < 0.01 vs Ctrl.shRNA group. (b) GSH production. RUNX1-specific shRNA or Ctrl.shRNA lentivirus-infected SHIN-3 and OVCAR-3 cells were treated with 2 or 10 μM erastin for 24 h. GSH production was measured as mentioned in Section 2. **p < 0.01 vs Ctrl.shRNA group. (c) MDA production. RUNX1-specific shRNA or Ctrl.shRNA lentivirus-infected SHIN-3 and OVCAR-3 cells were treated with 10 μM erastin for 24 h. **p < 0.01 vs Ctrl.shRNA group

Western blotting analysis. (a) SHIN-3 cells were infected with RUNX1 shRNA or control shRNA lentiviruses for 24 h. (b) The relative expression of RUNX1 was shown in histogram. **p < 0.01, compared with NC ShRNA

GO and KEGG analyses incorporating fold-change criteria

Ontology	ID	Description	Gene ratio	Bg Ratio	p value	p. adjust	z score
BP	GO:0002181	Cytoplasmic translation	20/189	146/18,800	2.72 × 10⁻¹⁷	9.34 × 10⁻¹⁴	−4.4721
BP	GO:0006979	Response to oxidative stress	21/189	433/18,800	2.83 × 10⁻⁹	4.86 × 10⁻⁶	2.4004
BP	GO:0000302	Response to reactive oxygen species	14/189	203/18,800	1.85 × 10⁻⁸	2.12 × 10⁻⁵	1.6036
BP	GO:0042060	Wound healing	19/189	429/18,800	7.1 × 10⁻⁸	6.09 × 10⁻⁵	2.5236
BP	GO:0034614	Cellular response to reactive oxygen species	11/189	147/18,800	2.86 × 10⁻⁷	0.0002	1.5076
CC	GO:0022626	Cytosolic ribosome	20/193	102/19,594	1.12 × 10⁻²⁰	3.65 × 10⁻¹⁸	−4.4721
CC	GO:0044391	Ribosomal subunit	20/193	178/19,594	9.76 × 10⁻¹⁶	1.59 × 10⁻¹³	−4.4721
CC	GO:0022627	Cytosolic small ribosomal subunit	12/193	43/19,594	6.94 × 10⁻¹⁵	7.54 × 10⁻¹³	−3.4641
CC	GO:0030055	Cell-substrate junction	27/193	428/19,594	1.62 × 10⁻¹⁴	1.32 × 10⁻¹²	0.57735
CC	GO:0005925	Focal adhesion	26/193	419/19,594	7.58 × 10⁻¹⁴	4.94 × 10⁻¹²	0.39223
MF	GO:0003735	Structural constituent of ribosome	20/190	181/18,410	3.18 × 10⁻¹⁵	1.41 × 10⁻¹²	−4.4721
MF	GO:0005200	Structural constituent of cytoskeleton	8/190	104/18,410	1.23 × 10⁻⁵	0.0027	1.4142
MF	GO:0019838	Growth factor binding	8/190	139/18,410	9.88 × 10⁻⁵	0.0123	1.4142
MF	GO:0038187	Pattern recognition receptor activity	4/190	26/18,410	0.0001	0.0123	2
MF	GO:0048027	mRNA 5′-UTR binding	4/190	27/18,410	0.0002	0.0123	−2
KEGG	hsa05171	Coronavirus disease – COVID-19	28/115	232/8,164	5.15 × 10⁻¹⁹	1.11 × 10⁻¹⁶	−2.2678
KEGG	hsa03010	Ribosome	20/115	158/8,164	3.91 × 10⁻¹⁴	4.2 × 10⁻¹²	−4.4721
KEGG	hsa04216	Ferroptosis	7/115	41/8,164	1.39 × 10⁻⁶	9.93 × 10⁻⁵	−0.37796
KEGG	hsa05144	Malaria	7/115	50/8164	5.55 × 10⁻⁶	0.0003	2.6458
KEGG	hsa04066	HIF-1 signaling pathway	8/115	109/8,164	0.0001	0.0061	0

Univariate and multivariate analysis

Characteristics	Total (N)	Univariate analysis		Multivariate analysis
Characteristics	Total (N)	HR (95% CI)	P value	HR (95% CI)	P value
FCGBP	379
Low	190	Reference		Reference
High	189	1.852 (1.425–2.406)	<0.001	1.524 (1.148–2.024)	0.004
ANKRD13A	379
Low	189	Reference		Reference
High	190	1.732 (1.334–2.248)	<0.001	1.352 (1.006–1.819)	0.046
TMEM181	379
Low	188	Reference		Reference
High	191	1.705 (1.314–2.211)	<0.001	1.216 (0.905–1.633)	0.195
RIPK4	379
Low	188	Reference		Reference
High	191	1.704 (1.310–2.215)	<0.001	1.115 (0.817–1.520)	0.493
RUNX1	379
Low	189	Reference		Reference
High	190	1.680 (1.293–2.182)	<0.001	1.429 (1.084–1.884)	0.011

Baseline characteristics

Characteristics	Low expression of RUNX1	High expression of RUNX1	P value
n	190	191
Race, n (%)			0.708
Asian	5 (1.4%)	7 (1.9%)
Black or African American	14 (3.8%)	11 (3%)
White	165 (45%)	165 (45%)
Age, n (%)			0.384
< = 60	100 (26.2%)	109 (28.6%)
>60	90 (23.6%)	82 (21.5%)
Clinical stage, n (%)			0.250
Stage I	1 (0.3%)	0 (0%)
Stage II	15 (4%)	8 (2.1%)
Stage III	146 (38.6%)	150 (39.7%)
Stage IV	26 (6.9%)	32 (8.5%)
Tumor status, n (%)			0.288
Tumor free	40 (11.8%)	32 (9.5%)
With tumor	129 (38.2%)	137 (40.5%)
Histologic grade, n (%)			1.000
G1	1 (0.3%)	0 (0%)
G2	22 (5.9%)	23 (6.2%)
G3 and G4	162 (43.7%)	163 (43.9%)
Primary therapy outcome, n (%)			0.069
PD	15 (4.9%)	12 (3.9%)
SD	10 (3.2%)	12 (3.9%)
PR	15 (4.9%)	28 (9.1%)
CR	122 (39.5%)	95 (30.7%)
Lymphatic invasion, n (%)			0.212
No	29 (19.5%)	19 (12.8%)
Yes	50 (33.6%)	51 (34.2%)
OS event, n (%)			0.004
Alive	87 (22.8%)	60 (15.7%)
Dead	103 (27%)	131 (34.4%)
DSS event, n (%)			0.019
No	89 (25.1%)	65 (18.3%)
Yes	91 (25.6%)	110 (31%)
PFI event, n (%)			0.933
No	51 (13.4%)	52 (13.6%)
Yes	139 (36.5%)	139 (36.5%)

Articles in this issue

DOI: https://doi.org/10.2478/ahem-2026-0003 | Journal eISSN: 1732-2693

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Language: English

Page range: 17 - 29

Submitted on: Aug 15, 2025

Accepted on: Feb 2, 2026

Published on: Mar 2, 2026

Published by: Hirszfeld Institute of Immunology and Experimental Therapy

In partnership with: Paradigm Publishing Services

Publication frequency: 1 issue per year

Keywords:

RUNX1,

Related subjects:

Microbiology and virology,

Medicine,

Basic medical science,

Immunology

© 2026 Deng He, Yu Zhou, Tiantian Feng, Hu Wang, Nenghuan Tang, Shangqi Ni, Wang Xi, Chengju Zhang, published by Hirszfeld Institute of Immunology and Experimental Therapy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Volume 80 (2026): Issue 1 (January 2026)

Diagnostic and prognostic value of RUNX1 in ovarian serous cystadenocarcinoma and its role in cancer progression via suppressing ferroptosis

Figures & Tables

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

GO and KEGG analyses incorporating fold-change criteria

Univariate and multivariate analysis

Baseline characteristics

Paradigm

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