Prospero homeobox 1 promotes proliferation, migration, and invasion of osteosarcoma cells and its clinical significance

Figures & data

Table 1. Primer sequences

Gene	Forward	Reverse
PROX1	CGAGAAGCCCAAACCAAATG	TGGTGGCTGCTTTGCTGTT
GAPDH	GTGGACCTGACCTGCCGTCT	GGAGGAGTGGGTGTCGCTGT
MMP28	TACCAACAGTTATGCGGCCTG	CAAAGCGTTTCTTACGCCTCA
HNF1A	CAGGTGATGAGCTACCAACC	AGGGTTCTTCTGCCTCTCAT
HNF1B	ACCAAGCCGGTCTTCCATACT	GGTGTGTCATAGTCGTCGCC
WNT2	TCCAGATGTGATGCGTGC	GAGTAGGACCCTGCCAAAA
CPZ	GCTTTGAGATCACGGTAGAGC	TCCACGAAATTCAGGAGTGACT
FZD9	CCACATCCGCAAGATCATGA	CATGTTGAGGCGTTCGTAGA
JAK3	CCTGATCGTGGTCCAGAGAG	GCAGGGATCTTGTGAAATGTCAT
MMP8	GGAACGCACTAACTTGACCTA	TGAGCATCTCCTCCAATACCT
β-catenin	GCAGTTCGCCTTCACTATGG	ATGTGCACGAACAAGCAACT

Table 2. Relationship between PROX1 expression and clinical characteristics of OS patients

Characteristic	Overall	Low expression of PROX1	High expression of PROX1	Chi-square test P-value
n	54	35	19
Gender				0.570
Male	34	23 (42.6)	11 (20.4)
Female	20	12 (22.2)	8 (14.8)
Age (years)				0.042
<21	43	25 (46.3)	18 (33.3)
≥21	11	10 (18.5)	1 (1.9)
Enneking stage				<0.001
IIA	4	4 (7.4)	0 (0)
IIB	28	24 (44.4)	4 (7.4)
IIIA	2	0 (0)	2 (10.5)
IIIB	20	7 (13.0)	13 (24.1)
M classification				<0.001
M0	32	28 (51.9)	4 (7.4)
M1	22	7 (13.0)	15 (27.8)
Histology				0.194
Conventional osteosarcoma	45	20 (37.0)	25 (46.3)
Others	9	2 (3.7)	7 (13.0)
Death				0.851
No	35	23 (42.6)	12 (22.2)
Yes	19	12 (22.2)	7 (13.0)
Tumor Size				0.724
<5 cm	10	6 (11.1)	4 (7.4)
≥5 cm	44	29 (53.7)	15 (24.8)

Conventional osteosarcoma: osteoblastic, chondroblastic and fibroblastic types;

Others: small cell, epithelioid, and telangiectatic types.

Figure 1. PROX1 expression was upregulated in OS tissues. (a) OS tissue with PROX1 high expression. (b) OS tissue with PROX1 moderately expression. (c) OS tissue with PROX1 weakly or negative expression. (d-e) The expression of PROX1 in normal bone tissue adjacent to the tumor and control OS group. OS, osteosarcoma.

Figure 2. High PROX1 expression predicted poor prognosis in OS patients. (a) The correlationship between PROX1 expression and metastasis in OS patients. (b) The overall survival of OS patients with high and low expression of PROX1. OS, osteosarcoma.

Figure 3. Effects of PROX1 knockdown on cell proliferation in OS cells. (a-b) qRT-PCR and Western blot analysis showed PROX1 expression in human osteoblast cell line (hFOB1.19) and four glioma cell lines (143B, U2OS, HOS and SAOS2). (c) The relative siPROX1 expression in 143B cell. (d) Proliferation in 143B cell was detected by CCK8 assay. OS, osteosarcoma. (*p < 0.05, **p < 0.01, ***p < 0.001).

Figure 4. Effects of PROX1 knockdown and overexpression on the migration and invasion of OS cells. (a) Wound healing assay was used to detect the cell migration ability after PROX1 knockdown or overexpression of 48 h in 143B cell. (b-c) Transwell assay showed the invasion of 143B cells after transfection with PROX1 siRNA or overexpression plasmid. OS, osteosarcoma.

Figure 5. The hierarchical aggregation analysis of DEGs in different samples. DEGs, differentially expressed genes.

Figure 6. The results of GO and KEGG enrichment analysis of DEGs. (a) The results of GO analysis of DEGs. GO, gene ontology; BP, biological process; CC, cellular component; MF, molecular function. (b) KEGG enrichment analysis results of DEGs. KEGG, Kyoto Encyclopedia of Genes and Genomes. DEGs, differentially expressed genes.

Figure 7. PPI network and important modules of DEGs. (a) The PPI network of DEGs. The red circles represent upregulated DEGs and blue circles represent downregulated DEGs. (b) The module 1 from the PPI network. (c) The module 2 from the PPI network. (d) The module 3 from the PPI network. PPI, protein-protein interaction; DEGs, differentially expressed genes.

Data availability statement

The data used to support the findings of this study are available from the corresponding author upon request.

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.