Survivin isoforms and clinicopathological characteristics in colorectal adenocarcinomas using real-time qPCR

Table 1 cDNA structure of survivin splice variants

Transcript variants	Exons or segments
Wild type survivin	1, 2, 3, 4
Survivin-2b	1, 2, 2b, 3, 4
SurvivinΔΕx3	1, 2, 4
Survivin-2a	1, 2
Survivin-3b	1, 2, 3, 3b, 4
Survivin-2b + 32	1, 2, 2b + 32nt, 3, 4

b: Sequence originated from intron.

Table 2 Clinical characteristics of 52 patients with colorectal cancer n (%)

Variables	Values
Gender
Men	21 (40.4)
Women	31 (59.6)
Tumor size
≤ 5 cm	32 (65.3)
> 5 cm	17 (34.7)
Grade
I	3 (6.3)
II	36 (75)
III	7 (14.6)
IV	2 (4.2)
Dukes stage
A	8 (16.3)
B	11 (22.4)
C	22 (44.9)
D	8 (16.3)
Morphologic cancer type
Polypoid (ecblastetic)	28 (57.1)
Ulcerous	21 (42.9)

Table 3 Sequences of primers and hybridization probes

	Name	Oligonucleotide sequence, 5’-3’
Forward primer	F1	CCACCGCATCTCTACATTCA
Reverse primers	WT	TATGTTCCTCTATGGGGTCG
	Sur2B	AGTGCTGGTATTACAGGCGT
	SurΔΕx3	TTTCCTTTGCATGGGGTC
Hybridization probes		CAAGTCTGGCTCGTTCTCAG TGGG-FITC1
		LCRed640-CAGTGGATGAAG CCAGCCTCG-Ph2

¹: 3’-end-labeled with fluorescein isothiocyanate (FITC);

²: 5’-end-labeled with LC Red 640; phosphorylated on the 3’ end to avoid extension of the probe.

Table 4 Quantitative measurements obtained from tissue mRNA and serum tumor markers from 52 patients with colorectal cancer (mean ± SD)

Variables	Values
Wild-survivin (copies/μL)	842.5 ± 2380.3
Survivin-2b (copies/μL)	58.1 ± 192.3
Survivin-ΔΕx3 (copies/μL)	393.9 ± 1988.8
Survivin-2b/wild-survivin (copies/μL)	0.12 ± 0.21
Survivin-ΔΕx3/wild-survivin (copies/μL)	0.36 ± 0.66
CEA (μg/L)	191.59 ± 934.62
CA 19-9 (μg/L)	892.48 ± 2991.78

CEA: Carcinoembryonic antigen; CA 19-9: Carbohydrate antigen 19-9.

Table 5 Spearman correlation coefficients of laboratory variables in patients with colorectal cancer (n = 52)

Variables	Sur2b	SurΔEx3	Sur2b/wildsur	SurΔEx3/wildsur	CEA	CA19-9	Age
Wildsur	0.694b	0.799b	0.408b	0.485b	0.136	0.102	0.051
Sur2b		0.623b	0.527b	0.471b	0.238	0.212	0.165
SurΔΕx3			0.409b	0.651b	0.208	0.063	0.229
Sur2b/wildsur				0.613b	0.021	0.203	0.0234
SurΔΕx3/wildsur					-0.054	0.084	0.320a
CEA						0.522b	0.395a
CA 19-9							0.247

^aP < 0.05,

^bP < 0.001. CEA: Carcinoembryonic antigen; CA 19-9: Carbohydrate antigen 19-9.

Table 6 Association of mRNA expression survivin splice variants with clinicopathological characteristics in 52 patients suffering from colorectal cancer (mean ± SD)

Clinical characteristics	n	Wild-sur (copies/μL)	P value wild-sur	Sur-2b (copies/μL)	P value sur-2b	Sur-ΔEx3 (copies/μL)	P value sur-ΔEx3	Sur-2b/wildsur (copies/μL)	P value sur-2b/wildsur	Sur-ΔEx3/wildsur (copies/μL)	P value sur-ΔEx3/wildsur
Gender			0.978		0.685		0.631		0.570		0.387
Male	20	1216.1 ± 3438.6		98.0 ± 299.7		834.8 ± 3085.6		0.08 ± 0.09		0.47 ± 0.39
Female	32	589.5 ± 1259.1		31.1 ± 33.2		85.3 ± 117.1		0.17 ± 0.24		0.36 ± 0,41
Tumor size			0.088		0.438		0.869		0.544		0.097
≤ 5 cm	32	308.4 ± 491.3		28.6 ± 34.6		96.2 ± 116.1		0.17 ± 0.24		0.48 ± 0.41
> 5 cm	17	1949.4 ± 3956.2		119.3 ± 330.9		1055.8 ± 3531.9		0.09 ± 0.09		0.23 ± 0.24
Dukes stage			0.469		0.737		0.544		0.351		0.097
A	8	212.4 ± 403.4		27.7 ± 27.7		51.7 ± 66.1		0.37 ± 0.40		0.44 ± 0.35
B	11	476.9 ± 743.4		30.8 ± 45.8		137.7 ± 185.7		0.08 ± 0.12		0.37 ± 0.37
C	22	1556.8 ± 3533.1		95.4 ± 292.5		782.6 ± 3019.7		0.09 ± 0.10		0.30 ± 0.39
D	8	226.7 ± 146.7		35.6 ± 32.9		120.7 ± 66.5		0.15 ± 0.15		0.63 ± 0.35
T			0.006		0.125		0.041		0.103		0.492
0-2	8	212.4 ± 403.4		27.7 ± 27.7		51.7 ± 66.1		0.37 ± 0.40		0.44 ± 0.35
3	29	340.0 ± 521.1		26.5 ± 35.4		92.9 ± 127.2		0.10 ± 0.13		0.39 ± 0.37
4	12	2620.8 ± 4579.4		162.9 ± 389.8		1413.2 ± 4056.1		0.10 ± 0.08		0.40 ± 0.44
Histologic type			0.584		0.030		0.731		0.061		0.892
Polypoid (ecblastetic)	28	520.1 ± 1006.4		45.7 ± 56.2		141.2 ± 313.7		0.19 ± 0.25		0.31 ± 0.25
Ulcerous	21	1354.5 ± 3550.4		79.3 ± 298.5		769.5 ± 3083.3		0.06 ± 0.06		0.54 ± 0.51