Overexpression of TLR3, TLR4, TLR7 and TLR9 in esophageal squamous cell carcinoma

Table 1 Polymerase chain reaction primers used for the detection of Toll-like receptors

TLR	Forward	Reverse
TLR3	AGTGCCGTCTATTTGCCACACA	AACAGTGCACTTGGTGGTGGAG
TLR4	TCTTCAACCAGACCTCTACATTCCA	GGAACATCCAGAGTGACATCACAG
TLR7	CCGTGACAATTACCTGGCCTTC	CAGGGCCTTCAGCTGGTTTC
TLR9	AGGATGATGCCAGGATGATGTC	TCAGGTCCAGGTTCTTGGTTGAG
β-actin	GGCACCCAGCACAATGAAG	CCGATCCACACGGAGTACTTG

TLR: Toll-like receptor.

Table 2 Statistical analysis of Toll-like receptor expression and clinicopathologic factors in esophageal carcinoma

Characteristics	TLR3	P	TLR4	P	TLR7	P	TLR9	P
	Positive (%)		Positive (%)		Positive (%)		Positive (%)
Normal control (n = 87)	7 (8)		5 (5.7)		8 (9.2)		4 (4.6)
Tumor differentiation	61 (70.1)		63 (72.4)		58 (66.7)		68 (78.2)
W (n = 30)	22 (75.3)	0.889	22 (75.3)	0.539	14 (46.7)	0.003	18 (60.0)	0.004
M (n = 29)	20 (69.1)		19 (65.6)		19 (65.6)		23 (79.3)
P (n = 28)	19 (67.9)		22 (78.6)		25 (89.3)		27 (96.4)
Depth of invasion
≤ muscularis (n = 37)	18 (48.6)	< 0.001	27 (73.1)	0.92	23 (62.2)	0.002	27 (73.1)	0.314
≥ adventitia (n = 50)	43 (86.0)		36 (72.0)		35 (66.0)		41 (82.0)
LN metastasis
Negative (n = 34)	18 (52.9)	0.005	20 (58.8)	0.023	26 (76.5)	0.12	29 (85.3)	0.197
Positive (n = 53)	43 (81.1)		43 (81.1)		32 (60.3)		39 (73.6)
Ethnic groups
Han (n = 30)	23 (76.7)	> 0.05	22 (73.3)	> 0.05	17 (56.7)	> 0.05	23 (76.7)	> 0.05
Uyghur (n = 25)	18 (72.0)		20 (80.0)		16 (64.0)		20 (80.0)
Kazak (n = 32)	20 (62.5)		21 (65.6)		25 (78.1)		25 (78.1)

All values are presented as the number of cases, with percentages in parentheses. W: Well differentiated; M: Moderately differentiated; P: Poorly differentiated; LN: Lymph node; TLR: Toll-like receptor.

Table 3 The percentage expression of Toll-like receptors in each cellular type within esophageal squamous cell carcinoma tissues

	Tumor cells	Fibroblast	MICs
Factors	Positive cases (%)	Positive cases (%)	Positive cases (%)
TLR3	61 (70.1)	3 (3.4)	4 (4.6)
TLR4	63 (72.4)	8 (9.2)	47 (48.3)
TLR7	58 (66.7)	5 (5.7)	2 (2.3)
TLR9	68 (78.2)	53 (60.9)	12 (13.8)

All values are presented as the number of cases, with percentages in parentheses. MICs: Mononuclear inflammatory cells; TLR: Toll-like receptor.

Table 4 Analysis of the relationship between the expression of Toll-like receptors in each cellular type with the clinicopathologic characteristics of esophageal squamous cell carcinoma n (%)

Factors	Tumor Grade			Depth of invasion		LN metastasis
	W	M	P	≤muscularis	≥adventitia	Negative	Positive
Number of cases	30	29	28	37	50	34	53
TLR4 MICs (+)	19 (63.3)	16 (55.2)	12(42.9)	12(32.4)	35(70)	14(41.2)	33(62.3)
P	0.245			0.001		0.018
TLR9 fibroblast (+)	20 (66.7)	18 (62.1)	17(60.7)	30(81.1)	23(43.4)	29(85.3)	24(45.3)
P	0.885			0.001		0.006

All values are presented as the number of cases, with percentages in parentheses. TLR: Toll-like receptor; LN: Lymph node; MICs: Mononuclear inflammatory cells; W: Well differentiated; M: Moderately differentiated; P: Poorly differentiated.