The clinical significance of the Ezrin gene and circulating tumor cells in osteosarcoma

Figures & data

Table 1 Characteristics of patients with osteosarcoma, n (%)

Variables	n	CanPatrol™
		Positive, n/total (%)	P-value
Age (years)
<18	26	25/26 (96.1)	0.261
≥18	15	13/15 (86.6)
Gender
Male	24	22/24 (91.7)	0.767
Female	17	16/17 (94.1)
Enneking stage
IA	1	0	<0.001
IB	2	0
IIA	1	1
IIB	26	26
IIIA	0	0
IIIB	11	11
Tumor location
Tibia or femur	33	31/33 (93.9)	0.53
Other	8	7/8 (87.5)
Tumor size
<5 cm	11	10	0.602
>5 cm	30	28

Figure 1 Fluorescence microscopy images of three types of CTCs isolated from the peripheral blood osteosarcoma patients, based on RNA-ISH staining of epithelial (red dots) and mesenchymal (green dots) markers.

Notes: (A) White blood cells, (B) epithelial CTCs, (C) mesenchymal CTCs, (D) epithelial/mesenchymal CTCs. Scale bar, 5 μm.
Abbreviations: CTCs, circulating tumor cells; ISH, in situ hybridization.

Table 2 Comparison of the CTC counts and the proportion of different CTC types between stage IIB and stage IIIB of patients with osteosarcoma

	IA, IB (n=3)	IIA	IIB (n=26)	IIIB (n=11)	P-value
CTC counts	0	8	24±6.74	80.72±14.78	0.00
Ratio of epithelial CTCs (%)	0	0	11.2±3.97	6.83±1.37	0.00
Ratio of mesenchymal CTCs (%)	0	0	31.92±5.35	70.00±8.27	0.00
Ratio of biophenotypic epithelial/mesenchymal CTCs (%)	0	0	56.87±6.83	23.15±4.43	0.00

Note: Data presented as mean ± standard deviation by calculating the CTC counts per patient in group IIB and IIIB using SPSS software 16.

Abbreviation: CTCs, circulating tumor cells.

Table 3 Different expression levels of Ezrin gene in CTCs of stage IIB and stage IIIB of patients with osteosarcoma

Enneking stage	CTC counts	High expression of Ezrin gene	Medium expression of Ezrin gene	Low expression of Ezrin gene	No expression of Ezrin gene
IIB	624	124	146	151	203
IIIB	888	423	212	121	132

Abbreviation: CTCs, circulating tumor cells.

Figure S1 Calibration curve obtained using the CanPatrol™ CTC enrichment technique in the spiking experiment (n=5) using 143B cells at different dilutions.

Abbreviation: CTCs, circulating tumor cells.

Table S1 The expression rates of the Ezrin gene in different types of CTCs

Types of CTCs	Positive for Ezrin gene (%)	Negative for Ezrin gene (%)
Epithelial CTCs	43.9	56.1
Mesenchymal CTCs	68.9	31.1
Biophenotypic epithelial/mesenchymal CTCs	81	19

Abbreviation: CTCs, circulating tumor cells.