Alcohol promotes epithelial mesenchymal transformation-mediated premetastatic niche formation of colorectal cancer by activating interaction between laminin-γ2 and integrin-β1

Table 1 Primer sequences for quantitative real-time polymerase chain reaction

Gene name	Forward (5’>3’)	Reverse (5’>3’)
For human
LAMC2	TTCTACAACGATCCGCACGAC	GAGTTAAAAGCAGCCCTGGT
ITGB1	CCTACTTCTGCACGATGTGATG	CCTTTGCTACGGTTGGTTACATT
FAK	GTCTGCCTTCGCTTCACG	GAATTTGTAACTGGAAGATGCAAG
Snail	CTAGCGAGTGGTTCTTCTGC	GTAGTTAGGCTTCCGATTGGG
E-cadherin	GACCGGTGCAATCTTCAAA	TTGACGCCGAGAGCTACAC
N-cadherin	AGCTTCTCACGGCATACACC	GTGCATGAAGGACAGCCTCT
Fibronectin	GCCTGGTACAGAATA TGTAGTG	ATCCCAGCTGATC AGTAGGCTGGTG
SATB1	GTGGAAGCCTTGGGAATCC	CTGACAGCTCTTCTTCTAGTT
GAPDH	GGAGCGAGATCCCTCCAAAAT	GGCTGTTGTCATACTTCTCATGG
For rats
LAMC2	GTGAAGCTTCCCTGCAAAAC	CATTTGGCCCCACGTAGT
ITGB1	TCGAAGGACCACTAGACCTGA	TTCCATGGGAACAAAAGGTAA
FAK	TTGGGTTGCAAACTATCTCTAGAAC	TGGTACAAAACTGGCACAGAA
Snail	TTCACATCCGAGTGGGTCTG	ACCCACACTGGTGAGAAGCC
E-cadherin	AAAGCAGGAAGAAAACACCACTC	AAAGGGCACGCTATCAACATTAG
N-cadherin	TCAGTGGCGGAGATCCTAC	GTGCTGAATTCCCTTGGCTA
Fibronectin	TGTCACCCACCACCTTGA	CTGATTGTTCTTCAGTGCGA
SATB1	AGATGCAGGGAGTGCCTTTA	TGCTCCTCCTTGCAATCATA
GAPDH	TGCCCTCATGTTCCTGATAAAT	CATTACATCACAGCTTTCCAGG

ITGB1: Integrin-β1; LAMC2: Laminin-γ2; SATB1: Special AT-rich sequence binding protein 1; FAK: Focal adhesion kinase; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Table 2 Number of rats with mucosal pathological tissue changes in different time periods

Wk	Group	n	Normal	Atypical hyperplasia	Adenoma	Adenocarcinoma		With lymphatic metastasis
						Mucosal carcinoma	Infiltrating carcinoma	Metastasis	Tumor metastasis rate
17	Non-alcohol	8	3	5	0	0	0	0	0.00%
	Alcohol	8	0	6	2	0	0	0	0.00%
24	Non-alcohol	8	0	0	0	6	2	0	0.00%
	Alcohol	8	0	0	0	2	6	3	37.5%

Table 3 Baseline characteristics of colorectal cancer patients

Baseline characteristics	Alcohol group (n = 29)	Non-alcohol group (n = 34)	P value
Gender
Male, n (%)	18 (62.1)	16 (47.1)	0.233
Female, n (%)	11 (37.9)	18 (52.9)
Age (yr old)
< 55, n (%)	10 (34.5)	14 (41.2)	0.586
≥ 55, n (%)	19 (65.5)	20 (58.8)
Differentiation
Medium, n (%)	24 (82.8)	30 (88.2)	0.536
Poor, n (%)	5 (17.2)	4 (11.8)
Clinical stage
Stage I-II, n (%)	12 (41.4)	24 (70.6)	0.020a
Stage III-IV, n (%)	17 (58.6)	10 (29.4)
T stage
T1-T2, n (%)	6 (20.7)	12 (35.3)	0.201
T3-T4, n (%)	23 (79.3)	22 (64.7)
Tumor metastasis conditions
Distant metastasis	12 (41.4)	23 (70.6)	0.046a
Lymphatic metastasis	9 (31)	8 (20.6)
Non-metastasis	8 (27.6)	3 (8.8)

^aP < 0.05.