Decreased expression of peroxiredoxin1 inhibits proliferation, invasion, and metastasis of ovarian cancer cell

Figures & data

Figure 1 The expression of PRDX1 in ovarian tissues (SP ×200, central SP ×400).

Notes: (A) Ovarian epithelial malignant tumor. (B) Ovarian epithelial borderline tumor. (C) Ovarian epithelial benign tumor. (D) Normal ovarian tissue. Scale bar: 300 µm.
Abbreviations: PRDX1, peroxiredoxin1; SP, scaled pixels.

Table 1 Expression of PRDX1 in ovarian tissues

Group	n	Low		High		Positive rate (%)	High positive rate (%)
		(−)	(+)	(++)	(+++)
Malignant	101	4	12	54	31	96.04*	84.16#
Borderline	11	3	5	3	0	72.72	27.27
Benign	21	9	9	2	1	57.14	14.29
Normal	15	12	3	0	0	20	0

Notes:

* Compared with the positive rate of borderline, benign, and normal groups, PRDX1 in the malignant group was higher (all P<0.01).

# Compared with the high positive rate of borderline, benign, and normal groups, PRDX1 in the malignant group was higher (all P<0.05).

Abbreviation: PRDX1, peroxiredoxin1.

Table 2 Relationship between PRDX1 expression and clinicopathologic parameters for malignant ovarian serous tumors

Item	n	Low		High		High positive rate (%)	P-value
		(−)	(+)	(++)	(+++)
FIGO stage							<0.05
I–II	39	4	8	17	10	69.23
III–IV	62	1	4	37	20	91.94
Differentiation
Well–moderate	44	3	11	21	9	55.00	<0.05
Poor	57	2	4	33	18	89.47
Lymph node metastasis							<0.05
Yes	29	0	3	18	8	89.66
No	65	4	16	26	19	69.23
Unknown*	7	0	1	6	0	85.71
Pathological subtype							>0.05
Serous	55	2	6	29	18	85.45
Mucinous	4	0	1	1	2	75.00
Endometrioid	14	0	3	7	4	70.57
Clear cell carcinoma	8	1	0	5	2	87.50
Poorly differentiated adenocarcinoma	20	1	2	12	5	85.00

Note:

* Patients without lymphadenectomy.

Abbreviations: FIGO, International Federation of Gynecology and Obstetrics; PRDX1, peroxiredoxin1.

Figure 2 Relationship between PRDX1 expression and clinicopathologic parameters.

Note: Correlation of PRDX1 expression with differentiation (A), stage (B), and lymph node metastasis (C).
Abbreviation: PRDX1, peroxiredoxin1.

Figure 3 Forest map based on univariate (A) and multivariate (B) Cox regression analyses.

Table 3 Cox regression analysis of overall survival of ovarian epithelial serous tumors

Variables	Univariate analysis			Multivariate analysis
	HR	95% CI of HR	P-value	HR	95% CI of HR	P-value
Age (<60 vs ≥60 years)	1.454	0.721–2.932	0.296	0.612	0.223–1.684	0.342
FIGO stage (I–II vs III–IV)	6.775	2.061–22.270	0.002*	4.569	1.226–17.031	0.024*
PRDX1 expression (low vs high)	7.154	2.171–23.570	0.001*	10.033	1.956–51.462	0.006*
Lymph node metastasis (yes vs no)	2.56	1.202–5.452	0.015*	0.737	0.294–1.845	0.514
Differentiation (poor vs well–moderate)	1.648	0.760–3.572	0.206	3.086	1.012–9.409	0.048*
Pathological subtype (serous vs nonserous)	1.230	0.604–2.502	0.568	2.787	0.794–9.783	0.110

Note:

* P<0.05.

Abbreviations: FIGO, International Federation of Gynecology and Obstetrics; PRDX1, peroxiredoxin1.

Note: *P<0.05.
Abbreviations: FIGO, International Federation of Gynecology and Obstetrics; PRDX1, peroxiredoxin1.

Table 3 Cox regression analysis of overall survival of ovarian epithelial serous tumors

Variables	Univariate analysis			Multivariate analysis
	HR	95% CI of HR	P-value	HR	95% CI of HR	P-value
Age (<60 vs ≥60 years)	1.454	0.721–2.932	0.296	0.612	0.223–1.684	0.342
FIGO stage (I–II vs III–IV)	6.775	2.061–22.270	0.002*	4.569	1.226–17.031	0.024*
PRDX1 expression (low vs high)	7.154	2.171–23.570	0.001*	10.033	1.956–51.462	0.006*
Lymph node metastasis (yes vs no)	2.56	1.202–5.452	0.015*	0.737	0.294–1.845	0.514
Differentiation (poor vs well–moderate)	1.648	0.760–3.572	0.206	3.086	1.012–9.409	0.048*
Pathological subtype (serous vs nonserous)	1.230	0.604–2.502	0.568	2.787	0.794–9.783	0.110

Note:

* P<0.05.

Abbreviations: FIGO, International Federation of Gynecology and Obstetrics; PRDX1, peroxiredoxin1.

Figure 4 Kaplan–Meier curves for overall survival of patients with epithelial ovarian cancer.

Note: Correlation of PRDX1 expression (A), clinical stage (B), and lymph node metastasis (C) with overall survival.
Abbreviations: Cum, cumulative; PRDX1, peroxiredoxin1.

Figure 5 Overall survival and expression differences of PRDX1 in different databases.

Notes: KM curves for overall survival of patients in KM plotter (A) and TCGA (B). Differential expression of PRDX1 in eight ovarian cancer data sets (C) and 11 cancer data sets (D) from Oncomine.
Abbreviations: PRDX1, peroxiredoxin1; KM, Kaplan–Meier; TCGA, the Cancer Genome Atlas; TPM, transcripts per million reads.

Figure 6 Correlation analysis of PRDX1 expression with copy number variation (A) and methylation data (B).

Abbreviations: PRDX1, peroxiredoxin1; FPKM, fragments per kilobase of transcript per million mapped reads.

Figure 7 Decreased PRDX1 inhibited proliferation, invasion, and metastasis capacities of ovarian cancer cells in vitro.

Notes: The expression of PRDX1 after PRDX1 knockdown by different siRNA was evaluated by immunoblot staining in ovarian cancer cell lines Caov-3 (A). Cell cycle of transfected cells was studied using flow cytometry (B). Invasion of PRDX1 knockdown ovarian cancer cells was measured by wound healing (C). Migration of PRDX1 knockdown cells was measured by transwell assay (D). The proliferating capability of transfected cells was evaluated using MTT (E). Quantification of the immunoblot staining results (a), flow cytometry results (b), wound healing assay results (c), and transwell invasion assay results (d). siRNA1, siRNA2, and siRNA3 denote PRDX1-siRNA transfection. NC indicates an empty-plasmid transfected cell line and CA is an untreated cell line. *P<0.05, **P<0.01.
Abbreviations: PRDX1, peroxiredoxin1; siRNA, small interfering RNA.

Figure 8 PRDX1 correlated enrichment gene analysis.

Notes: (A) Wnt signaling pathway (P=0.0028; FDR=0.077; enrichment score=−0.349). (B) Oxidative phosphorylation (P=0.000; FDR=0.000; enrichment score=0.739). (C) Pathways in cancer (P=0; FDR=0.058; enrichment score=−0.325). (D) Immunoblot staining results showed the expression levels of E-CA, VIM, β-catenin after gene transfection. (E) Quantification of the immunoblot staining results. (F) The change of cell morphology, before and after PRDX1 gene transfection. NC indicates an empty-plasmid transfected cell line and CA is an untreated cell line. **P<0.05.
Abbreviations: PRDX1, peroxiredoxin1; FDR, false discovery rate; siRNA, small interfering RNA; E-CA, E-cadherin; VIM, vimentin; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Figure 9 Network of enriched terms and top 30 of GO terms and pathway enrichment analysis.

Notes: (A) Bubble pattern of enriched biological function of GO terms in the genes associated with PRDX1. (B) Bubble pattern of enriched pathway in the genes associated with PRDX1. The Y-axis represents the name of the pathway or function, and the X-axis represents the percentage of this node in the whole network. The size of the bubble represents the number of genes in this signaling pathway or the number of genes involved in this function. The color of the bubble indicates the P-value of enrichment. The darker the color, the more significant the result.
Abbreviations: PRDX1, peroxiredoxin1; GO, Gene Ontology.

Figure 10 PRDX1 protein–protein interaction network.

Notes: Each node represented a protein, and the edge between two nodes represented the relationship between two proteins. Different thicknesses of an edge represented the strength of the interaction. The more interaction the protein had, the bigger the node.
Abbreviations: PRDX1, peroxiredoxin1; MCODE, Molecular Complex Detection.

Figure 11 The MCODE module of PPI and biological function and pathways analysis.

Notes: (A) MCODE1. Each node represented a protein and the edge between two nodes represented the relationship between two proteins. (B) Network of pathways of MCODE1. The circle represents the regulatory gene and the diamond represents the pathway. (C) Bar graph of function enrichment of genes in MCODE1. The Y-axis represents a signaling pathway or function. The X-axis represents the number of genes enriched in this region. The bar color indicates the statistical significance. The bluer the bar, the smaller the P-value.
Abbreviations: MCODE, Molecular Complex Detection; GO, Gene Ontology; PPI, protein–protein interaction.

Figure 12 Flowchart of the bioinformatics approach.

Note: Chart of functional enrichment analysis of PRDX1-related genes and construction of protein interaction network.
Abbreviations: PRDX1, peroxiredoxin1; MCODE, Molecular Complex Detection; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.