Mucinous phenotype and CD10 expression of primary adenocarcinoma of the small intestine

Table 1 Primers used in this study

		Primer
KRAS	Forward	5’-GGCCTGCTGAAAATGACTGA-3’
	Reverse	5’-GTTGGATCATATTCGTCCAC-3’
BRAF	Forward	5’-CCTTTACTTACTACACCTCAG-3’
	Reverse	5’-CATCCACAAAATGGATCCAG-3’
BAT25	Forward1	5’-TCGCCTCCAAGAATGTAAGT-3’
	Reverse	5’-TCTGCATTTTAACTATGGCTC-3’
BAT26	Forward1	5’-TGACTACTTTTGACTTCAGCC-3’
	Reverse	5’-AACCATTCAACATTTTTAACCC-3’
D2S123	Forward1	5’-GAAACAGGATCCTGCCTTTA-3’
	Reverse	5’-GGACTTTCCACCTATGGGAC-3’
D5S346	Forward1	5’-GACTCACTCTAGTATAAATCGGG-3’
	Reverse	5’-GAGCAGATAAGACAGTATTACTAGTT-3’
D17S250	Forward1	5’-GGGAAGAATCAAATAGACAAT-3’
	Reverse	5’-GCTGGCCATATATATATTTAAACC-3’

¹Forward primers for microsatellite instability (MSI) analysis were fluorescently-labeled with 6-carboxyfluorescein (6-FAM).

Table 2 Clinicopathological findings of all 47 cases of small intestinal adenocarcinomas n (%)

	All cases (n = 47)
Age (yr)
Mean (range)	59.9 (23-87)1
Gender
Male	29
Female	18
Gross appearance
Polypoid	12
Ulcerative	35
Size (cm) (mean)	5.13
Location
Duodenum	14
Jejunum	21
Ileum	12
Histological subtype
Tubular2	33
Poorly	9
Mucinous	5
Invasion depth3
T1	2
T2	4
T3	18
T4	23
Lymphatic permeation	33 (70)
Venous invasion	12 (26)
Adenomatous component	3 (6)

¹Age of one patient was unknown;

²"Tubular" contains well or moderately differentiated Adenocarcinoma;

³UICC TNM classification is used.

Table 3 Clinicopathological features and expression of mucin core proteins and CD10

	MUC5AC		MUC6		MUC2		CD10
	Positive(n = 12)	Negative(n = 35)	Positive(n = 4)	Negative(n = 43)	Positive(n = 20)	Negative(n = 27)	Positive(n = 7)	Negative(n = 40)
Age (yr)
Mean ± SD	63.4 ± 15.11	58.8 ± 16.5	76.3 ± 10.01	58.7 ± 15.9	62.3 ± 15.3	58.0 ± 16.71	60.1 ± 15.1	59.8 ± 16.51
(range)	(35-85)	(23-87)	(67-87)	(23-85)	(38-87)	(23-81)	(38-87)	(23-85)
Gender (male/female)	10/2	19/16	3/1	26/17	9/11	18/9	3/4	27/14
Gross appearance
(polypoid/ulcerated)	3/9	9/26	2/2	10/33	6/14	6/21	4/3a	8/32a
Size (cm)	5.3 ± 2.2	5.1 ± 1.9	5.0 ± 1.8	5.1 ± 2.0	5.7 ± 1.6a	4.7 ± 2.2a	5.9 ± 1.4a	5.0 ± 2.1a
Location
(duodenum/jejunum/ileum)	2/5/5	12/16/7	2/1/1	12/20/11	6/11/3	8/10/9	4/2/1	10/19/11
Histological subtype
(tubular2/poorly/mucinous)	6/3/3	27/6/2	3/0/1	30/9/4	13/2/5	10/7/0	7/0/0	29/9/5
Invasion depth3
T1	0	2	0	2	2	0	2a	0a
T2	2	2	2	2	2	2	2a	2a
T3	2	16	0	18	6	12	1a	17a
T4	8	15	2	21	10	13	2a	21a
Lymphatic permeation n (%)	10 (83)	23 (66)	2 (50)	31 (72)	12 (60)	21 (78)	3 (43)	30 (75)
Venous invasion n (%)	3 (25)	9 (26)	1 (25)	11 (26)	5 (25)	7 (26)	2 (29)	10 (25)
Adenomatous component n (%)	0 (0)	3 (9)	1 (25)	2 (5)	3 (15)a	0 (0)a	2 (29)a	1 (3)a

¹Age of one patient was unknown;

²"tubular" contains well or moderately differentiated adenocarcinoma;

³UICC TNM classification is used.

^aP < 0.05 vs positive group and negative group in the same mucin core protein or CD10. In comparison invasion depth, T1 vs T2-T4).

Table 4 Summarized data of microsatellite instability status, expression of mismatch repair proteins and KRAS mutations

	MSI	Expression of MMR protein		KRAS mutationbase exchange (codon)	Age	Gender
		MLH1	MSH2
Case 6	MSI-H	+	+	-	55	M
Case 36	MSI-H	+	-	-	85	M
Case 31	MSI-H	-	+	-	67	M
Case 34	MSI-H	+	+	-	71	M
Case 12	MSI-L	+	+	GGT>GAT(12)	69	M
Case 28	MSI-L	+	-	-	41	M
Case 24	MSI-L	+	+	-	60	M
Case 40	MSI-L	+	+	-	35	M
Case 25	MSI-L	+	+	-	53	F
Case 5	MSI-L	+	+	-	23	F
Case 44	MSI-L	+	+	-	87	F
Case 38	MSI-L	+	+	-	67	F
Case 41	MSS	+	+	GGC>GAC(13)	45	F
Case 20	MSS	+	+	GGT>GAT(12)	59	F
Case 46	MSS	+	+	GGT>GTT(12)	40	F

MSI: Microsatellite instability; MSI-H: MSI-high; MSI-L: MSI-low; MMR: Mismatch repair; M: Male; F: Female.

Table 5 Clinicopathological features, microsatellite instability status and KRAS mutation

	MSI		KRAS
	MSI-H(n = 4)	MSI-L/MSS(n = 30)	Mutational type(n = 4)	Wild type(n = 33)
Age (yr)	69.5 ± 12.3	59.6 ± 15.91	53.3 ± 13.2	60.8 ± 16.21
Gender (male/female)	4/0	16/14	1/3	22/11
Gross appearance (polypoid/ulcerated)	1/3	6/24	2/2	9/24
Size (cm)	7.7 ± 1.1a	5.1 ± 2.0a	4.4 ± 1.0	5.5 ± 2.0
Location (duodenum/jejunum/ileum)	0/2/2	7/15/8	1/2/1	9/15/9
Histological subtype (tubular2/poorly/mucinous)	2/0/2	22/6/2	4/0/0	22/8/3
Invasion depth3
T1	0	1	0	1
T2	0	2	0	3
T3	2	13	1	13
T4	2	14	3	15
Lymphatic permeation n (%)	3 (75)	23 (77)	4 (100)	22 (67)
Venous invasion n (%)	1 (25)	9 (30)	1 (25)	10 (30)
Adenomatous component n (%)	0 (0)	1 (3)	0 (0)	2 (6)

¹Age of one patient was unknown;

²"Tubular" contains well or moderately differentiated adenocarcinoma;

³UICC TNM classification is used.

^aP < 0.05 vs control.

Table 6 Summarized data of 4 small intestinal adenocarcinomas with co-expression of MUC5AC and MUC2

	Age	Gender	Location	Histological subtype	Invasion depth	Lymphatic permeation	Venous invasion	Lymph node metastasis	MSI
case 6	55	m	Jejunum	Tubular	T3	-	-	-	MSI-H
case 29	61	m	Jejunum	Mucinous	T4	+	-	No data	MSS
case 31	67	m	Ileum	Mucinous	T4	+	-	No data	MSI-H
case 36	85	m	Ileum	Mucinous	T4	+	-	-	MSI-H

MSI: Microsatellite instability; MSI-H: MSI-high; MSS: Microsatellite stable; M: Male.