Underexpression of LATS1 TSG in colorectal cancer is associated with promoter hypermethylation

Table 1 Clinical and histopathological characteristics of colorectal cancer patients and results of large tumor suppressor 1 mRNA quantification using quantitative polymerase chain reaction n (%)

Clinical parameter			Blood parameters (mean ± SD)				MSI results (n = 84)					QPCR results
			RBC (106/μL)	Ht	Hb (g/dL)	WBC (103/μL)	BAT 26	BAT 25	BAT 40	MSI-L	MSI-H	Downregulated cases vs control
	CRC 142 cases		4.26 ± 4.58	36% ± 5.2%	12 ± 2.1	7.41 ± 2.71	28/84 (33)	25/84 (30)	26/84 (31)	30/84 (36)	14/84 (17)	127/142 (88)1
Sex and age (mean ± SD, range, yr)	M (n = 87)	67 ± 10.4 (37-89)	4.36 ± 0.502	36% ± 6.0%	12 ± 2.4	7.26 ± 2.91	21/54 (39)	18/54 (33)	16/54 (30)	22/54 (41)	9/54 (17)	78/87 (90)
	F (n = 55)	69 ± 11.4 (44-90)	4.18 ± 0.41	35% ± 4.6%	11 ± 1.8	7.53 ± 2.58	7/30 (23)	7/30 (23)	10/30 (33)	8/30 (37)	5/30 (17)	49/55 (88)
Tumor location	Right side	55 (39)	4.09 ± 0.462	33% ± 4.7%2	10 ± 1.92	7.18 ± 3.45	9/32 (28)	8/32 (25)	7/32 (22)	8/32 (25)	4/32 (12.5)	51/55 (93)
	Left side	87 (61)	4.36 ± 0.432	38% ± 4.8%2	12 ± 1.82	7.56 ± 2.17	19/52 (36.5)	17/52 (33)	19/52 (36.5)	22/52 (42)	10/52 (19)	76/87 (81)
	Ascending colon	46 (32)	4.08 ± 0.46	33% ± 5.0%2	11 ± 2.02	7.26 ± 3.42	9/26 (35)	8/26 (31)	6/26 (23)	7/26 (27)	4/26 (15)	43/46 (93)1
	Transverse colon	11 (8)	4.12 ± 0.43	33% ± 4.2%2	10 ± 1.52	7.43 ± 3.47	0/6 (0)	0/6 (0)	1/6 (17)	1/6 (17)	0/6 (0)	10/11 (91)1
	Descending/sigmoid colon	41 (29)	4.29 ± 0.53	36% ± 5.2%2	12 ± 2.12	7.49 ± 2.35	6/23 (26)	7/23 (30)	9/23 (39)	9/23 (39)	4/23 (17)	32/41 (78)1
	Rectum	44 (31)	4.43 ± 0.33	39% ± 4.5%2	13 ± 1.62	7.47 ± 2.08	13/29 (45)	10/29 (34)	10/29 (34)	13/29 (45)	6/29 (21)	42/44 (95)1
Dukes’ stage	A	27 (19)	4.23 ± 0.49	38% ± 5.3%	12 ± 2.0	7.58 ± 2.08	2/19 (11)	2/19 (11)2	1/19 (6)2	4/19 (22)	0/19 (0)	22/27 (81)12
	B	41 (29)	4.30 ± 0.47	36% ± 5.5%	11 ± 2.1	7.38 ± 1.99	9/23 (39)	5/23 (22)2	7/23 (30)2	8/23 (35)	4/23 (17)	40/41 (98)1
	C	54 (38)	4.22 ± 0.47	36% ± 5.0%	12 ± 2.0	7.42 ± 3.41	12/34 (35)	14/34 (41)2	15/34 (44)2	14/34 (41)	8/34 (24)	49/54 (91)1
	D	20 (14)	4.34 ± 0.26	32% ± 4.4%	10 ± 1.9	7.13 ± 2.26	5/8 (62.5)	4/8 (50)2	3/8 (37.5)2	4/8 (50)	2/8 (25)	18/22 (82)1
Lymph node metastasis	Negative	68 (48)	4.28 ± 0.46	36% ± 5.5%	12 ± 2.1	7.44 ± 1.97	11/42 (26)	7/42 (17)2	8/42 (19)2	12/42 (29)2	4/42 (10)2	62/68 (91)1
	Positive	74 (52)	4.23 ± 0.45	35% ± 5.0%	11 ± 2.1	7.39 ± 3.28	17/42 (40)	18/42 (43)2	18/42 (43)2	18/42 (43)2	10/42 (24)2	65/74 (88)1
Histological differentiation (G stage)	Well (G1)	3 (2)	4.17 ± 0.35	36% ± 4.7%	12 ± 1.4	7.44 ± 2.19	1/3 (33)	1/3 (33)	1/3 (33)	0/3 (0)	1/3 (33)	3/3 (100)1
	Moderate (G2)	48 (34)	4.26 ± 0.47	36% ± 5.0%	11 ± 2.0	7.17 ± 2.29	8/31 (26)	6/31 (19)	8/31 (26)	9/31 (29)	4/31 (13)	46/48 (96)1
	Poorly (G3)	88 (62)	4.16 ± 0.45	35% ± 5.5%	11 ± 2.2	7.62 ± 3.54	18/47 (38)	18/47 (38)	17/47 (36)	20/47 (43)	9/47 (19)	84/88 (95)1
	Undifferentiated (G4)	3 (2)	4.36 ± 0.79	37% ± 6.6%	12 ± 2.3	9.16 ± 4.74	1/3 (33)	0/3 (0)	0/3 (0)	1/3 (33)	0/3 (0)	2/3 (66)1

¹Statistically significant difference between analyzed subgroup and control;

²Statistically significant difference between subgroups, Kruskal-Wallis analysis of variance test. RBC: Red blood cells count; Ht: Hematocrit; Hb: Hemoglobin; WBC: White blood cells; MSI: Microsatellite instability; BAT26: Marker for MSH2; BAT25: Marker for the c-kit oncogene; BAT40: Marker for the HSD3B2; MSI-L: Low grade MSI, mutation in 1-2 BAT markers; MSI-H: High grade MSI, mutation in 3 BAT markers; CRC: Colorectal cancer; M: Male; F: Female.

Table 2 Relationship between large tumor suppressor 1 promoter methylation status, large tumor suppressor 1 mRNA level, and histopathological and microsatellite instability data in colorectal cancer n (%)

Clinical parameter		Total	M	Av. LATS1 mRNA fold	UM	Av. LATS1 mRNA fold	Av. LATS1 mRNA fold	P value between
				change, control vs M		change, control vs UM	change, UM vs M	UM and M groups
Tumor total		44	25 (57)	597	19	3.55	162	0.00005
Dukes’ stage	A	4	4 (100)	556	0	No data	No data	-
	B	16	5 (31)	699	11	228	3	0.041
	C	19	11 (58)	469	8	1.53	305	0.009
	D	5	5 (100)	1263	0	No data	No data	-
Lymph node metastasis	Negative	20	9 (45)	632	11	75	8	0.015
	Positive	24	16 (67)	586	8	1.53	381	0.0002
Histological differentiation G stage	G1	2	1 (50)	801	1	538	1.5	NS
	G2	11	3 (27)	1216	8	1.5	802	0.018
	G3	28	20 (71)	538	8	166	3.65	0.015
	G4	3	1 (33)	699	2	92	8	NS
MSI status	MSS	27	19 (70)	7	8	1.7	4	NS
	MSI-L	11	7 (64)	8	4	2.4	3.3	NS
	MSI-H	6	4 (67)	12	2	3.5	3.4	NS

M: Hypermethylation of large tumor suppressor 1 (LATS1) promoter; UM: Unmethylation of LATS1 promoter; MSI: Microsatellite instability; MSS: Microsatellite stable; MSI-L: Low grade MSI, mutation in 1-2 BAT markers; MSI-H: High grade MSI, mutation in 3 BAT markers; NS: Noy significant.