Upregulation of STC2 in colorectal cancer and its clinicopathological significance

Figures & data

Figure 1 Increase of STC2 expression in GEO and TCGA databases in CRC.

Notes: (A) DEGs in CRC vs normal tissues across each independent dataset. Each column represents a sample and each row represents the expression level of a given mRNA. The color scale represents the raw Z score ranging from blue (low expression) to red (high expression). (B) Venn diagrams of overlapping DEGs between GSE21510, GSE32323, GSE39582, and GSE41328. (C) Analysis of STC2 expression in unpaired CRC (N=349) and normal tissues (N=41) in TCGA (P<0.001). (D) Analysis of STC2 expression in CRC and PANT (N=41) in TCGA (P<0.001). (E) High STC2 expression was associated with poor OS in CRC patients in TCGA.

Abbreviations: CRC, colorectal cancer; DEGs, differently expressed genes; GEO, Gene Expression Omnibus; OS, overall survival; PANT, paired adjacent normal tissues; STC2, stanniocalcin 2; TCGA, The Cancer Genome Atlas.

Figure 2 High expression of STC2 validated by our own database in CRC.

Notes: (A) DEGs in six pairs of CRC and PANT. (B) STC2 protein and mRNA levels were measured in CRC and PANT by Western blotting and qRT-PCR, respectively.

Abbreviations: CRC, colorectal cancer; DEGs, differently expressed genes; PANT, paired adjacent normal tissues; qRT-PCR, quantitative real-time polymerase chain reaction; STC2, stanniocalcin 2.

Table 1 Univariate and multivariate analyses of clinicopathological characteristics and STC2 with overall survival in TCGA

	Univariate analysis		Multivariate analysis
	HR (95% CI)	P-value	HR (95% CI)	P-value
Gender (male vs female)	0.520 (0.264–1.022)	0.058
Age, years (≥median vs <median)	1.756 (0.903–3.413)	0.097
Vascular invasion (present vs absent)	2.459 (1.280–4.725)	0.007	1.275 (0.472–3.448)	0.632
Lymphatic invasion (present vs absent)	2.361 (1.228–4.537)	0.010	1.327 (0.486–3.627)	0.581
Tumor infiltration (T3 and T4 vs T1 and T2)	2.060 (0.489–8.673)	0.324
Lymph node metastasis (present vs absent)	1.743 (0.911–3.333)	0.093
Node counts (≥12 vs <12)	0.486 (0.227–1.042)	0.064
Distant metastasis (present vs absent)	3.562 (1.804–7.031)	<0.001	2.444 (1.004–5.950)	0.049
Clinical stage (III and IV vs I and II)	2.237 (1.147–4.361)	0.018	1.174 (0.482–2.861)	0.725
STC2 (≥median vs <median)	2.338 (1.189–4.599)	0.014	2.064 (1.035–4.113)	0.040

Note: Bold indicates statistically significant values (P<0.05).

Abbreviations: STC2, stanniocalcin 2; TCGA, The Cancer Genome Atlas.

Figure 3 Detection of STC2 protein expression in tissues by IHC staining, and high STC2 expression in tumors predicted poor prognosis in CRC patients.

Notes: (A) Representative images of STC2 protein expression in CRC and PANT. STC2 protein expression was significantly increased in tumor specimens compared with PANT. (B) IHC staining of STC2 protein in CRC tissues. The staining intensity was scored as 0 (negative), 1 (weak positive), 2 (moderated positive), and 3 (strong positive). (C) High expression of STC2 had a worse OS for CRC patients.

Abbreviations: CRC, colorectal cancer; IHC, immunohistochemistry; OS, overall survival; PANT, paired adjacent normal tissues; STC2, stanniocalcin 2.

Table 2 Correlations between STC2 expression level and clinicopathological variables of 115 CRC patients

	Number	STC2 expression level		P-value
		Low (n=63)	High (n=52)
Gender				0.595
Male	61	32	29
Female	54	31	23
Age, years				0.932
<60	57	31	26
≥60	58	32	26
Tumor location				0.136
Colon	62	30	32
Rectum	53	33	20
Tumor size, cm				0.105
<5	47	30	17
≥5	68	33	35
Serum CEA, ng/mL				0.904
<5	56	31	25
≥5	59	32	27
Vascular invasion				0.655
Present	33	17	16
Absent	82	46	36
Lymphatic invasion				0.137
Present	34	15	19
Absent	81	48	33
Tumor infiltration				0.205
T1 and T2	38	24	14
T3 and T4	77	39	38
Lymph node metastasis				0.047
Present	46	20	26
Absent	69	43	26
Node counts				0.055
<12	38	16	22
≥12	77	47	30
Distant metastasis				0.040
Present	17	5	12
Absent	98	58	40
Clinical stage				0.047
I	19	14	5
II	45	27	18
III	34	17	17
IV	17	5	12

Note: Bold indicates statistically significant values (P<0.05).

Abbreviations: CEA, carcinoembryonic antigen; CRC, colorectal cancer; STC2, stanniocalcin 2.

Table 3 Univariate and multivariate analyses of clinicopathological characteristics and STC2 with overall survival in 115 CRC patients

	Univariate analysis		Multivariate analysis
	HR (95% CI)	P-value	HR (95% CI)	P-value
Gender (male vs female)	1.155 (0.659–2.022)	0.615
Age, years (≥median vs <median)	0.749 (0.427–1.313)	0.313
Tumor location (rectum vs colon)	0.840 (0.478–1.475)	0.682
Tumor size, cm (≥5 vs <5)	1.730 (0.949–3.153)	0.074
Serum CEA, ng/mL (≥5 vs <5)	0.842 (0.481–1.474)	0.546
Vascular invasion (present vs absent)	2.311 (1.302–4.100)	0.004	1.875 (0.992–3.545)	0.053
Lymphatic invasion (present vs absent)	1.876 (1.054–3.341)	0.033	1.740 (0.940–3.221)	0.078
Tumor infiltration (T3 and T4 vs T1 and T2)	1.626 (0.861–3.072)	0.112
Lymph node metastasis (present vs absent)	2.391 (1.337–4.275)	0.003	1.303 (0.391–4.339)	0.666
Node counts (≥12 vs <12)	1.364 (0.733–2.537)	0.327
Distant metastasis (present vs absent)	5.057 (2.647–9.662)	<0.001	3.742 (1.633–8.575)	0.002
Clinical stage (III and IV vs I and II)	3.015 (1.650–5.508)	<0.001	1.335 (0.344–5.176)	0.676
STC2 (≥median vs <median)	2.273 (1.278–4.042)	0.005	1.976 (1.092–3.576)	0.024

Note: Bold indicates statistically significant values (P<0.05).

Abbreviations: CEA, carcinoembryonic antigen; CRC, colorectal cancer; STC2, stanniocalcin 2.

Figure S1 Differentially expressed genes in CRC vs normal tissues across each independent dataset. Each column represents a sample and each row represents the expression level of a given mRNA. The color scale represents the raw Z score ranging from blue (low expression) to red (high expression).

Abbreviation: CRC, colorectal cancer.

Table S1 Microarray datasets of mRNA expression profiles

Dataset	Year	Country	Organization	Platform	Sample		PMID
					Normal tissue	CRC tissue
GSE21510^Citation1	2011	Japan	Juntendo University	GPL570	25	19	21270110
GSE32323^Citation2	2012	Japan	Juntendo University	GPL570	17	17	22399497
GSE39582^Citation3	2013	France	Ligue Nationale Contre le Cancer	GPL570	17	17	23700391
GSE41328^Citation4	2012	USA	Illinois University	GPL570	10	10	17160039

Abbreviation: CRC, colorectal cancer.

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