Upregulation of nemo-like kinase is an independent prognostic factor in colorectal cancer

Table 1 Differential expression of nemo-like kinase protein in normal mucosa, adenoma, and various stages of colorectal cancer tissue specimens

Characteristic	Total	NLK expression				Positive rate	Mean rank1	P value2
		-	+	++	+++
Normal mucosa	56	14	35	6	1	75.0%	216.32	-
Adenoma	51	7	31	13	0	86.3%	273.63	0.049a
Stage I	53	4	22	21	6	92.5%	392.97	0.005c
Stage II	312	31	97	154	30	90.1%	410.66	0.573
Stage III	298	24	76	165	33	91.9%	441.42	0.074
Stage IV	49	1	7	17	24	98.0%	596.47	< 0.001b

¹Kruskal–Wallis test was used for the overall comparison (P < 0.001);

²Mann-Whitney U test was used to determine significant differences between two groups.

^aP≤ 0.05 vs normal mucosa;

^cP≤ 0.05 vs adenoma;

^bP≤ 0.01 vs stage III cancer.

Table 2 Association of nemo-like kinase expression with clinicopathologic parameters of colorectal cancer patients (n = 712)

Parameter	NLK immunostaining				P value
	-	+	++	+++
Sex					0.1651
Male	29	112	210	55
Female	31	90	147	38
Age (yr)					0.4081
< 60	33	129	211	65
≥ 60	27	73	146	28
Tumor location					< 0.0011
Colon cancer	48	110	145	36
Rectal cancer	12	92	212	57
Invasion depth					0.9801
T1-T2	6	27	42	12
T3-T4	54	175	315	81
Lymph node metastasis					0.0272
N0	35	122	180	43
N1	21	54	120	36
N2	4	26	57	14
Distant metastasis					< 0.0011
M0	59	195	340	69
M1	1	7	17	24
TNM stage					< 0.0012
I	4	22	21	6
II	31	97	154	30
III	24	76	165	33
IV	1	7	17	24
Tumor differentiation					0.0191
Well, moderate	42	182	326	82
Poor, mucinous	18	20	31	11
Serum CEA (ng/mL)					0.5701
< 5	39	123	220	63
≥ 5	21	79	137	30
Serum CA19-9 (U/mL)					0.2641
< 37	51	178	294	79
≥ 37	9	24	63	14
Recurrence					< 0.0011
No	56	176	257	68
Yes	4	26	80	25

¹Mann-Whitney U test;

²Kruskal-Wallis test. CA19-9: Cancer antigen 19-9; CEA: Carcinoembryonic antigen; TNM: Tumor-node-metastasis.

Table 3 Univariate and multivariate analyses in colorectal cancer tissue specimens (n = 712)

Variables	Overall survival		Disease-free survival
	HR (95%CI)	P value	HR (95%CI)	P value
Univariate analyses
Sex (male vs female)	0.84 (0.58-1.23)	0.376	0.86 (0.63-1.18)	0.342
Age (≥ 60 yr vs < 60 yr)	1.24 (0.85-1.80)	0.259	0.95 (0.69-1.30)	0.734
TNM stage (IV/III vs II/I)	3.11 (2.06-4.70)	< 0.001	2.37 (1.72-3.27)	< 0.001
Differentiation (poor/mucinous adenocarcinoma vs well/moderate)	1.54 (0.93-2.55)	0.094	1.33 (0.85-2.08)	0.219
Position (rectal vs colon)	0.56 (0.38-0.83)	0.003	0.92 (0.66-1.23)	0.506
CEA (≥ 5 ng/mL vs < 5 ng/mL)	1.97 (1.36-2.85)	< 0.001	1.63 (1.19-2.21)	0.002
CA19-9 (≥ 37 U/mL vs < 37 U/mL)	2.27 (1.50-3.42)	< 0.001	1.89 (1.31-2.72)	0.001
NLK (++/+++ vs -/+)	2.19 (1.44-3.35)	< 0.001	2.71 (1.86-3.94)	< 0.001
Postoperative radiochemotherapy (yes vs no)	2.42 (1.30-4.51)	0.005	2.69 (1.60-4.50)	< 0.001
Multivariate analyses
TNM stage (IV/III vs II/I)	3.02 (1.99-4.59)	< 0.001	2.34 (1.70-3.22)	< 0.001
Differentiation (poor/mucinous adenocarcinoma vs well/moderate)	1.78 (1.06-2.98)	0.029	1.45 (0.94-2.24)	0.091
Position (rectal vs colon)	0.49 (0.33-0.72)	< 0.001	0.80 (0.59-1.08)	0.143
CEA (≥ 5 ng/mL vs < 5 ng/mL)	1.56 (1.04-2.35)	0.032	1.31 (0.95-1.81)	0.099
CA19-9 (≥ 37 U/mL vs < 37 U/mL)	1.54 (0.99-2.41)	0.058	1.76 (1.24-2.49)	0.002
NLK (++/+++ vs -/+)	2.57 (1.66-3.98)	< 0.001	1.96 (1.40-2.74)	< 0.001
Postoperative radiochemotherapy (yes vs no)	1.04 (0.50-2.16)	0.921	1.64 (0.93-2.92)	0.090

CA19-9: Cancer antigen 19-9; CEA: Carcinoembryonic antigen; CI: Confidence interval; HR: Hazard ratio; NLK: Nemo-like kinase; TNM: Tumor-node-metastasis.