Connective tissue growth factor expression hints at aggressive nature of colorectal cancer

Table 1 Clinicoepidemological and clinicopathological parameters of study population (n = 71)

Characteristics	n (%)
Age in years
< 50	21 (29.58)
≥ 50	50 (70.42)
Gender
Male	38 (53.52)
Female	33 (46.48)
Dwelling
Rural	47 (66.20)
Urban	24 (33.80)
Social class	22 (30.99)
Low
Middle and high	49 (69.01)
Family history
Yes	20 (28.17)
No	51 (71.83)
Smoking status
Yes	40 (56.34)
No	31(43.66)
Lifestyle
Active	31 (43.66)
Sedentary	40 (56.34)
Salt tea intake
Yes	65 (91.55)
No	06 (8.45)
Red meat consumption
Yes	59 (83.10)
No	12 (16.90)
Sundried vegetables
Yes	48 (67.61)
No	23 (32.39)
Source of drinking water
Tap water (R)	46 (64.79)
Tap water (L)	07 (9.86)
Others1	18 (25.35)
Pickles
Yes	41 (57.75)
No	30 (42.25)
Pesticide exposure
Yes	33 (46.48)
No	38 (53.52)
Junk food consumption
Yes	05 (7.04)
No	66 (92.96)
Site of tumor
Colon	36 (50.70)
Rectum	20 (28.17)
Rectosigmoid	15 (21.13)
Tumor differentiation
Well	18 (25.35)
Moderate	46 (64.79)
Poor	07 (9.86)
Invasion depth
T1	08 (11.27)
T2	22 (30.99)
T3	31 (43.66)
T4	10 (14.08)
T1 + T2	30 (42.25)
T3 + T4	41 (57.75)
TNM staging
I	25 (35.21)
II	25 (35.21)
III	18 (25.35)
IV	03 (4.22)
I + II	50 (70.42)
III + IV	21 (29.58)
Tumor grade
1	18 (25.35)
2	46 (64.79)
3	07 (9.86)
DUKE stage
A	05 (7.04)
B	44 (61.97)
C	22 (30.99)
Node status
0	51 (71.83)
1 and 2	20 (28.17)
LVI
Present	54 (76.06)
Absent	17 (23.94)
PNI
Present	15 (21.12)
Absent	56 (78.87)
TALNR
Present	62 (87.32)
Absent	09 (12.68)
Necrosis seen
Yes	18 (25.35)
No	53 (74.65)
Recurrence
Yes	12 (16.90)
No	59 (83.10)
Vital status
Alive	66 (92.96)
Dead	05 (7.04)

¹Others: Spring, Well, Pond.

N: Total Number of samples; R: River water through Tap; L: Lake water through Tap; TNM: Tumor-node-metastasis; TALNR: Tumor associated lymph-node response; LVI: Lymphovascular invasion; PNI: Perineural invasion.

Table 2 Clinicopathological relevance of connective tissue growth factor mRNA expression in colorectal cancer patients as determined by quantitative real-time polymerase chain reaction

Characteristics	Normal expression (n = 14)	Overexpression (n = 57)	Odds ratio (95%CI)	P value	Chi2
Age
< 50	2 (9.52)	19 (90.48)	Referent
≥ 50	12 (24.00)	38 (76.00)	3.0 (0.6-29.9)	0.16	1.95
Gender
Male	4 (10.53)	34 (89.47)	Referent
Female	10 (30.30)	23 (69.70)	3.6 (0.9-17.8)	0.037a	4.36
Dwelling
Rural	11 (23.40)	36 (76.60)	Referent
Urban	3 (12.50)	21 (87.50)	0.5 (0.1-2.1)	0.27	1.19
Social class
Low	7 (31.82)	15 (68.18)	Referent
Middle and high	7 (14.29)	42 (85.71)	0.4 (0.09-1.4)	0.08	2.94
Family history
Yes	3 (15.00)	17 (85.00)	Referent
No	11 (21.57)	40 (78.43)	1.5 (0.3-9.7)	0.53	0.39
Smoking status
Yes	4 (10.00)	36 (90.00)	Referent
No	10 (32.26)	21 (67.74)	4.3 (1.04-20.68)	0.019a	5.46
Lifestyle
Active	8 (25.81)	23 (74.19)	Referent
Sedentary	6 (15.00)	34 (85.00)	0.5 (0.1-1.1)	0.25	1.28
Salt tea intake
Yes	13 (20.00)	52 (80.00)	Referent
No	1 (16.67)	5 (83.33)	0.8 (0.02-8.1)	0.844	0.03
Red meat consumption
Yes	13 (22.03)	46 (77.97)	Referent
No	1 (8.33)	11 (91.67)	0.3 (0.01-2.6)	0.37	1.22
Sundried vegetables
Yes	9 (18.75)	39 (81.25)	Referent
No	5 (21.74)	18 (78.26)	1.2 (0.2-4.7)	0.767	0.08
Source of drinking water
Tap water (R)	12 (26.09)	34 (73.91)	Referent
Tap water (L)	1 (14.29)	6 (85.71)	0.5 (0.01-5.3)
Others1	1 (5.56)	17 (94.44)	0.4 (0.04-2.4)	0.54	1.21
Pickles
Yes	8 (19.51)	33 (80.49)	Referent
No	6 (20.00)	24 (80.00)	1.03 (0.3-3.9)	1.000	0.002
Pesticide exposure
Yes	7 (21.21)	26 (78.79)	Referent
No	7 (18.42)	31 (81.58)	0.84 (0.219-3.21)	0.8	0.07
Junk food consumption
Yes	0 (0)	5 (100)	Referent
No	14 (21.21)	52 (91.23)	0 (0-3.08)	0.25	1.32
Site of tumor
Colon	7 (19.44)	29 (80.56)	Referent
Rectum	6 (30.00)	14 (70.00)	1.8 (0.4-7.45)
Rectosigmoid	1 (6.67)	14 (93.33)	0.3 (0.01-2.7)	0.23	2.91
Tumor differentiation
Well	8 (44.44)	10 (55.56)
Moderate	5 (10.87)	41 (89.13)	Referent
Poor	1 (14.29)	6 (85.71)	0.15 (0.03-0.7)	0.009a	9.35
Invasion depth
T1	5 (62.50)	3 (37.50)
T2	7 (31.82)	15 (68.18)	Referent
T3	2 (6.45)	29 (93.55)	0.3 (0.03-2.0)	0.001a	17.21
T4	0 (0)	10 (100)
T1 + T2	12 (40.00)	18 (60.00)
T3 + T4	2 (4.88)	39 (95.12)	0.08 (0.008-0.4)	0.000a	13.5
TNM staging
I	11 (44.00)	14 (56.00)	Referent
II	2 (8.00)	23 (92.00)	0.1 (0.01-0.6)	0.002a	14.5
III	1 (5.56)	17 (94.44)	0.07 (0.01-0.7)
IV	0 (0)	3 (100.00)		0.040a	4.21
I + II	13 (26.00)	37 (74.00)
III + IV	1 (4.76)	20 (95.24)	0.1 (0.01-1.1)
Tumor grade
1	8 (44.44)	10 (55.56)
2	5 (10.87)	41 (89.13)	Referent
3	1 (14.29)	6 (85.71)	0.1 (0.03-0.7)	0.009a	9.35
DUKE stage
A	2 (2.82)	3 (4.23)
B	11 (15.49)	33 (46.48)	Referent
C	1 (1.41)	21 (29.58)	0.5 (0.05-6.8)	0.042a	5.20
Node status
0	13 (25.49)	38 (74.51)	Referent
1 and 2	1 (5.00)	19 (95.00)	0.1 (0.003-1.2)	0.041a	3.81
LVI
Present	8 (14.81)	46 (85.19)	Referent
Absent	6 (35.29)	11 (64.71)	3.1 (0.7-12.7)	0.064	3.42
PNI
Present	0 (0.00)	15 (100)	Referent
Absent	14 (25.00)	42 (75.00)	0 (0-1.08)	0.031a	4.67
TALNR
Present	11 (17.74)	51 (82.26)	Referent
Absent	3 (33.33)	6 (66.67)	2.3 (0.32-12.8)	0.272	1.20
Necrosis seen
Yes	17 (94.44)	1 (5.56)	Referent
No	40 (75.47)	13 (24.53)	2.7 (0.5-27.7)	0.080	3.10
Recurrence
Yes	0 (0)	12 (100)	Referent
No	14 (23.73)	45 (76.27)	0 (0-1.08)	0.060	3.55
Vital status
Alive	13 (19.70)	53 (80.30)	Referent
Death	1 (20.00)	4 (80.00)	1.4 (0.02-1.9)	0.85	0.07

¹Others: Spring, Well, Pond.

^aP < 0.05: Statistical significance.

R: River water through tap; L: Lake water through tap; TALNR: Tumor associated lymph-node response; LVI: Lymphovascular invasion; PNI: Perineural invasion; CI: Confidence interval.

Table 3 Clinicopathological relevance of connective tissue growth factor protein expression in colorectal cancer patients as determined by Western blot analysis

Characteristics	Normal expression (n = 28)	Overexpression (n = 43)	Odds ratio (95%CI)	P value	Chi2
Age
< 50	6 (28.57)	15 (71.43)	Referent
≥ 50	22 (44.00)	28 (56.00)	1.9 (0.6-7.1)	0.22	1.47
Gender
Male	12 (31.58)	26 (68.42)	Referent
Female	16 (48.48)	17 (51.52)	2.03 (0.7-6.0)	0.15	2.11
Dwelling
Rural	20 (42.55)	27 (57.45)	Referent
Urban	8 (33.33)	16 (66.67)	0.7 (0.21-2.1)	0.45	0.57
Social class
Low	10 (45.45)	12 (54.55)	Referent
Middle and high	18 (36.73)	31 (63.27)	1.4 (0.5-4.5)	0.57	0.48
Family history
Yes	7 (35.00)	13 (65.00)	Referent
No	21 (41.18)	30 (58.82)	0.7 (0.2-2.5)	0.63	0.23
Smoking status
Yes	11 (27.50)	29 (72.50)	Referent
No	17 (54.84)	14 (45.16)	3.2 (1.1-9.7)	0.019a	5.46
Lifestyle
Active	12 (38.71)	19 (61.29)	Referent
Sedentary	16 (40.00)	24 (60.00)	1.05 (0.4-0.1)	0.91	0.01
Salt tea intake
Yes	25 (38.46)	40 (61.54)	Referent
No	3 (50)	3 (50)	1.6 (0.2-12.8)	0.58	0.30
Red meat consumption
Yes	24 (40.68)	35 (59.32)	Referent
No	4 (33.33)	8 (66.67)	0.7 (0.1-3.1)	0.63	0.22
Sundried vegetables
Yes	19 (39.58)	29 (60.42)	Referent
No	9 (39.13)	14 (60.87)	1.0 (0.3-3.0)	0.97	0.01
Source of drinking water
Tap water (R)	20 (43.48)	26 (56.52)	Referent
Tap water (L)	1 (14.29)	6 (85.71)	0.21 (0.01-2.3)
Others1	7 (38.89)	11 (61.11)	1.3 (0.3-4.6)	0.34	2.21
Pickles
Yes	16 (39.02)	25 (60.98)	Referent
No	12 (40)	18 (60)	1.04 (0.3-3.03)	0.93	0.01
Pesticide exposure
Yes	15 (45.45)	18 (54.55)	Referent
No	13 (34.21)	25 (65.79)	0.624 (0.2-1.8)	0.33	0.93
Junk food consumption
Yes	0 (0)	5 (100)	Referent
No	28 (42.42)	38 (57.58)	0 (0-1.1)	0.06	3.5
Site of tumor
Colon	12 (33.33)	24 (66.67)	Referent
Rectum	12 (60)	8 (40)	3 (0.84-10.89)
Rectosigmoid	4 (26.67)	11 (73.33)	0.7 (0.13-3.1)	0.077	5.12
Tumor differentiation
Well	14 (77.78)	4 (22.22)	Referent
Moderate	13 (28.26)	33 (71.74)	0.1 (0.02-0.5)
Poor	1 (14.29)	6 (85.71)	0.04 (0.01-0.64)	0.000a	15.33
Invasion depth
T1	7 (87.50)	1 (12.50)
T2	13 (59.09)	9 (40.91)	Referent	0.000a	18.61
T3	7 (22.58)	24 (77.42)	0.04 (0.01-0.44)
T4	1 (10)	9 (90)
T1 + T2	20 (66.67)	10 (33.33)
T3 + T4	8 (19.51)	33 (80.49)	0.12 (0.03-0.4)	0.000a	16.12
TNM staging
I	19 (76.00)	6 (24.00)	Referent
II	7 (28.00)	18 (72.00)	0.12 (0.02-0.5)	0.000a	23.3
III	2 (11.11)	16 (88.89)	0.04 (0.003-0.3)
IV	0 (0.00)	3 (100)	0 (0-0.5)
I + II	26 (52.00)	24 (48.00)
III + IV	2 (9.52)	19 (90.48)	0.1 (0.01-0.5)	0.001a	11.2
Tumor grade
1	14 (77.78)	4 (22.22)	Referent
2	13 (28.26)	33 (71.74)	0.1 (0.02-0.5)
3	1 (14.29)	6 (85.71)	0.05 (0.01-0.6)	0.000a	15.33
DUKE stage
A	4 (80.00)	1 (20.00)	Referent
B	22 (50.00)	22 (50.00)	4 (0.3-205)
C	2 (9.09)	20 (90.91)	0.02 (0.01-0.5)	0.001a	13.9
Node status
0	26 (50.98)	25 (49.02)	Referent
1 and 2	2 (7.14)	18 (41.86)	0.10 (0.01-0.53)	0.001a	10.10
LVI
Present	17 (31.48)	37 (68.52)	Referent
Absent	11 (64.71)	6 (35.29)	0.2 (0.06-0.90)	0.015a	5.97
PNI
Present	1 (6.67)	14 (93.33)	Referent
Absent	27 (48.21)	29 (51.79)	0.1 (0.01-0.58)	0.003a	8.55
TALNR
Present	25 (40.32)	37 (59.68)	Referent
Absent	3 (33.33)	6 (66.67)	0.7 (0.11-3.9)	0.688	0.16
Necrosis seen
Yes	2 (11.11)	16 (88.89)	Referent
No	26 (49.06)	27 (50.94)	0.2 (0.03-0.76)	0.004a	8.10
Recurrence
Yes	1 (8.33)	11 (91.67)	Referent
No	27 (45.76)	32 (54.24)	0.3 (0.05-1.34)	0.016a	5.85
Vital status
Alive	27 (40.91)	39 (59.09)	Referent
Death	1 (20.00)	4 (80.00)	0.4 (0.007-3.9)	0.36	0.85

¹Others: Spring, Well, Pond.

^aP < 0.05: Statistical significance.

R: River water through tap, L: Lake water through tap; TALNR: Tumor associated lymph-node response; LVI: Lymphovascular invasion; PNI: Perineural invasion; CI: Confidence interval.

Table 4 Clinicopathological relevance of connective tissue growth factor high and low expression status determined by immunohistochemistry in colorectal cancer patients

Characteristics	Low expression (n = 27)	High expression (n = 44)	Odds ratio (95%CI)	P value	Chi2
Age
< 50	6 (28.57)	15 (71.43)	Referent
≥ 50	21 (42.00)	29 (58.00)	1.8 (0.5-6.6)	0.28	1.13
Gender
Male	11 (28.95)	27 (71.05)	Referent
Female	16 (48.48)	17 (51.52)	2.3 (0.8-6.9)	0.09	2.86
Dwelling
Rural	19 (40.43)	28 (59.57)	Referent
Urban	8 (33.33)	16 (66.67)	0.7 (0.2-2.2)	0.56	0.33
Social class
Low	10 (45.45)	12 (54.55)	Referent
Middle and high	17 (34.69)	32 (65.31)	1.5 (0.5-5.0)	0.38	0.74
Family history
Yes	7 (35.00)	13 (65.00)	Referent
No	20 (39.22)	31 (60.78)	0.8 (0.2-2.7)	0.74	0.10
Smoking status
Yes	10 (25.00)	30 (75.00)	Referent
No	17 (54.84)	14 (45.16)	3.6 (1.2-11.3)	0.010a	6.61
Lifestyle
Active	12 (38.71)	19 (61.29)	Referent
Sedentary	15 (37.50)	25 (62.50)	0.9 (0.3-2.8)	0.92	0.01
Salt tea intake
Yes	24 (36.92)	41 (63.08)	Referent
No	3 (50.00)	3 (50.00)	1.7 (0.2-13.6)	0.53	0.41
Red meat consumption
Yes	23 (38.98)	36 (61.02)	Referent
No	4 (33.33)	8 (66.67)	0.8 (0.15-3.3)	0.71	0.13
Sundried vegetables
Yes	19 (39.58)	29 (60.42)	Referent
No	8 (34.78)	15 (65.22)	0.8 (0.2-2.5)	0.70	0.15
Source of drinking water
Tap water (R)	20 (43.48)	26 (56.52)	Referent
Tap water (L)	1 (14.29)	6 (85.71)	0.2 (0.004-2.06)
Others1	6 (33.33)	12 (66.67)	0.6 (0.2-2.3)	0.29	2.41
Pickles
Yes	16 (59.26)	25 (56.82)	Referent
No	11 (40.74)	19 (43.18)	0.9 (0.3-2.6)	0.84	0.04
Pesticide exposure
Yes	15 (55.56)	18 (40.91)	Referent
No	12 (44.44)	26 (59.09)	0.5 (0.2-1.6)	0.23	1.4
Junk food consumption
Yes	0 (0)	5 (100)	Referent
No	27 (40.91)	39 (59.09)	0 (0-1.1)	0.07	3.30
Site of tumor
Colon	11 (30.56)	25 (69.44)	Referent
Rectum	12 (60.00)	8 (40.00)	3.4 (0.9-12.5)
Rectosigmoid	4 (26.67)	11 (73.33)	0.8 (0.1-3.7)	0.06	5.81
Tumor differentiation
Well	14 (77.78)	4 (22.22)	Referent
Moderate	12 (26.09)	34 (73.91)	0.1 (0.02-0.4)
Poor	1 (14.29)	6 (85.71)	0.05 (0.01-0.6)	0.000a	16.52
Invasion depth
T1	7 (25.93)	1 (2.27)	Referent
T2	12 (44.44)	10 (22.73)	0.2 (0.003-1.8)	0.001a	17.30
T3	7 (25.93)	24 (54.550)	0.04 (0.01-0.4)
T4	1 (3.70)	9 (20.45)	0.01 (0.01-0.40)
T1 + T2	19 (70.37)	11 (25.00)
T3 + T4	8 (29.63)	33 (75.00)	0.14 (0.04-0.4)	0.000a	14.11
TNM staging
I	18 (66.67)	7 (15.91)	Referent
II	7 (25.93)	18 (40.91)	0.1 (0.03-0.6)	0.000a	20.7
III	2 (7.41)	16 (36.36)	0.05 (0.01-0.3)
IV	0 (0)	3 (6.820)	0 (0-0.6)
I + II	25 (92.59)	25 (56.82)
III + IV	2 (7.41)	19 (43.18)	0.12 (0.01-0.5)	0.001a	10.31
Tumor grade
1	14 (77.78)	4 (22.22)	Referent
2	12 (26.09)	34 (73.91)	0.1 (0.02-0.4)
3	1 (14.29)	6 (85.71)	0.05 (0.01-0.6)	0.000a	16.52
DUKE stage
A	4 (80.00)	1 (20.00)	Referent
B	21 (47.73)	23 (52.27)	0.2 (0.004-2.6)
C	2 (9.09)	20 (90.91)	0.02 (0.014-0.5)	0.001a	13.3
Node status
0	25 (49.02)	26 (50.98)	Referent
1 and 2	2 (10.00)	18 (90.00)	0.1 (0.01-0.6)	0.002a	9.30
LVI
Present	17 (31.48)	37 (68.52)	Referent
Absent	10 (58.82)	7 (41.18)	0.3 (0.1-1.13)	0.043a	4.11
PNI
Present	1 (6.67)	14 (93.33)	Referent
Absent	26 (46.43)	30 (53.57)	0.1 (0.002-0.6)	0.020a	5.42
TALNR
Present	24 (38.71)	38 (61.29)	Referent
Absent	3 (33.33)	6 (66.67)	0.8 (0.1-4.1)	0.75	0.09
Necrosis seen
Yes	24 (47.06)	27 (52.94)	Referent
No	3 (15.00)	17 (85.00)	0.2 (0.03-0.82)	0.012a	6.21
Recurrence
Yes	1 (8.33)	11 (91.67)	Referent
No	26 (44.07)	33 (55.93)	0.11 (0.012-0.9)	0.020a	5.40
Vital status
Alive	26 (39.39)	40 (60.61)	Referent
Death	1 (20.00)	4 (80.00)	0.4 (0.07-4.2)	0.389	0.74

¹Others: Spring, Well, Pond.

^aP < 0.05: Statistical significance.

R: River water through tap; L: Lake water through tap; TALNR: Tumor associated lymph-node response; LVI: Lymphovascular invasion; PNI: Perineural invasion; CI: Confidence interval.

Table 5 Predictors for recurrence or mortality of colorectal cancer using extended cox-regression analysis model

Characteristics	Disease-free survival			Overall survival
	HR	95%CI	P value	HR	95%CI	P value
Age	1.66	0.4-6.4	0.45	1.03	0.9-1.1	0.32
Tumor differentiation	1.13	0.5-2.7	0.79	3.1	0.7-14.4	0.15
Tumor grade	1.12	0.4- 2.7	0.79	3.1	0.7-14.4	0.15
Depth invasion	1.6	0.8-3.2	0.18	1.91	0.6-5.7	0.30
TNM stage	2.8	1.38-5.97	0.005a	3.7	1.1-12.7	0.043a
Duke stage	9.6	2.3-40.5	0.002a	3.36	0.6-18.3	0.161
Node status	0.08	0.02-0.37	0.001a	0.25	0.04-1.5	0.139
Necrosis	0.26	0.1-0.82	0.022a	1.30	0.14-11.6	0.811
LVI	0.25	0.05-1.2	0.090	1.16	3.25e-17	0.08
PNI	0.47	0.14-1.6	0.230	0.16	0.02-0.98	0.041a
CTGF expression	0.13	0.01-1.06	0.013a	0.39	0.04-3.5	0.33

^aP < 0.05: Statistical significance. HR: Hazard ratio; CI: Confidence interval; TNM: Tumor-node-metastasis; LVI: Lymphovascular invasion; PNI: Perineural invasion; CTGF: Connective tissue growth factor.