Relationship between Fusobacterium nucleatum, inflammatory mediators and microRNAs in colorectal carcinogenesis

Table 1 Relative quantification of mRNA expression of the inflammatory genes in adenoma and colorectal cancer samples compared with adjacent normal tissue samples and colorectal cancer samples relative to the adenoma group

	TLR2	TLR4	NFKB1	IL1B	IL6	IL8	TNF
CRA
RQ median	0.87	2.27	1.02	2.27	1.12	3.33	0.75
RQ Range	0.36-10.80	0.32-11.25	0.45-3.38	0.14-76.76	0.09-80.41	0.10-874.50	0.14-2.60
P value	0.2940	0.0003a	0.5360	0.0047a	0.2473	0.0006a	0.0527
CRC
RQ median	2.36	0.74	0.90	4.13	6.67	6.36	0.72
RQ Range	0.31-35.80	0.20-23.42	0.31-8.49	0.13-245.70	0.09-192.80	0.16-194.20	0.06-31.76
P value	< 0.0001a	0.4476	0.9855	< 0.0001a	< 0.0001a	< 0.0001a	0.2391
CRC/CRA
RQ median	1.68	0.54	0.78	4.79	9.40	12.14	0.70
RQ Range	0.22-25.49	0.15-17.22	0.27-7.35	0.15-284.80	0.13-271.80	0.31-370.60	0.06-30.77
P value	< 0.0001a	0.0608	0.1237	< 0.0001a	< 0.0001a	< 0.0001a	0.1560

Wilcoxon signed rank test.

^aP value < 0.05 were considered statistically significant. CRA: Colorectal adenoma; CRC: Colorectal cancer; RQ: Relative quantification; TNF: Tumour necrosis factor; IL: Interleukin; NFKB: Nuclear factor kappa B; TLR: Toll-like receptor.

Table 2 Relative quantification of microRNAs in adenoma and colorectal cancer samples compared with adjacent normal tissue samples and colorectal cancer samples relative to the adenoma group

	miR-21	miR-22	miR-28	miR-34a	miR-135b
CRA
RQ median	0.75	0.85	0.73	0.97	2.19
RQ Range	0.15-13.91	0.10-3.05	0.08-2.17	0.12-4.74	0.10-25.13
P value	0.6349	0.1747	0.1904	0.3306	0.0074a
CRC
RQ median	0.53	0.27	0.65	1.38	9.31
RQ Range	0.08-12.53	0.026-2.78	0.11-7.27	0.20-30.0	0.45-74.00
P value	0.1725	< 0.0001a	0.0045a	0.0029a	< 0.0001a
CRC/CRA
RQ median	1.01	0.59	0.77	1.26	2.64
RQ Range	0.15-23.91	0.06-6.07	0.14-8.79	0.18-27.51	0.13-20.96
P value	0.0969	0.5770	0.1271	0.0101a	< 0.0001a

Wilcoxon signed rank test.

^aP value < 0.05 were considered statistically significant. CRA: Colorectal adenoma; CRC: Colorectal cancer; RQ: Relative quantification.

Table 3 Correlation analysis between the relative quantification of the inflammatory genes and microRNAs with the quantification of Fusobacterium nucleatum in adenoma and colorectal cancer samples

Spearman correlation coefficient (r)	CRA	CRC
TLR2	0.42	0.09
P value	0.1557	0.6335
TLR4	-0.62	-0.01
P value	0.0235a	0.9587
NFKB1	-0.40	-0.04
P value	0.1809	0.8045
IL1B	0.31	0.46
P value	0.3064	0.0066a
IL6	0.01	0.47
P value	0.9716	0.0059a
IL8	-0.07	0.54
P value	0.8166	0.0013a
TNF	0.30	0.23
P value	0.3156	0.2027
miR-21	-0.21	0.26
P value	0.4643	0.1467
miR-22	0.17	0.38
P value	0.5528	0.0331a
miR-28	0.04	0.01
P value	0.8872	0.9413
miR-34a	0.17	0.00
P value	0.5630	0.9905
miR-135b	-0.05	0.22
P value	0.8637	0.2163

Spearman correlation test.

^aP value < 0.05 were considered statistically significant. CRA: Colorectal adenoma; CRC: Colorectal cancer; TNF: Tumour necrosis factor; IL: Interleukin; NFKB: Nuclear factor kappa B; TLR: Toll-like receptor.

Table 4 Correlation analysis between the relative quantification of microRNAs with the inflammatory genes in adenoma and colorectal cancer samples

Spearman correlation coefficient (r)	miR-21	miR-22	miR-28	miR-34a	miR-135b
CRA
TLR2	0.09	0.29	0.31	0.23	0.09
P value	0.6608	0.1655	0.1266	0.2606	0.6714
TLR4	0.18	0.06	0.18	0.41	0.38
P value	0.3892	0.7926	0.3955	0.0431	0.0598
NFKB1	-0.01	-0.17	-0.21	-0.19	-0.20
P value	0.9796	0.4252	0.3191	0.3650	0.3454
IL1B	0.29	0.15	0.09	0.45	0.38
P value	0.1667	0.4834	0.6555	0.0232	0.0610
IL6	0.30	0.08	0.05	0.26	0.24
P value	0.1430	0.7148	0.8237	0.2136	0.2417
IL8	0.40	0.18	0.17	0.44	0.46
P value	0.0466a	0.4017	0.4273	0.0296a	0.0200a
TNF	-0.21	-0.04	-0.03	-0.05	-0.11
P value	0.3083	0.8638	0.9012	0.8152	0.6084
CRC
TLR2	0.29	0.53	0.18	0.19	-0.14
P value	0.0777	0.0005a	0.2790	0.2408	0.3923
TLR4	0.07	0.34	0.33	0.26	0.05
P value	0.668	0.0377a	0.0401a	0.1216	0.7586
NFKB1	0.18	0.31	0.02	-0.10	-0.30
P value	0.2779	0.0499a	0.8958	0.5462	0.0574
IL1B	0.53	0.43	0.13	0.22	-0.10
P value	0.0004a	0.0055a	0.4139	0.1668	0.5248
IL6	0.54	0.57	0.18	0.21	0.05
P value	0.0004a	0.0002a	0.2813	0.2095	0.7680
IL8	0.53	0.37	0.09	0.23	0.13
P value	0.0005a	0.0180a	0.5734	0.1506	0.4314
TNF	0.26	0.42	0.03	0.08	-0.32
P value	0.1094	0.0071a	0.8406	0.6264	0.0411a

Spearman correlation test.

^aP value < 0.05 were considered statistically significant. CRA: Colorectal adenoma; CRC: Colorectal cancer; TNF: Tumour necrosis factor; IL: Interleukin; NFKB: Nuclear factor kappa B; TLR: Toll-like receptor.

Table 5 KRAS mutation and microsatellite instability status in adenoma and colorectal cancer groups

KRAS status/mutation type	CRA, n = 27 (%)	CRC, n = 43 (%)	P value
WT	19 (70.4)	32 (74.5)	0.7854
Mutation	8	11
p.Gly12Ala (G12A)	1 (3.7)	0
p.Gly12Ser (G12S)	1 (3.7)	1 (2.3)
p.Gly12Val (G12V)	1 (3.7)	2 (4.6)
p.Gly12Asp (G12D)	3 (11.1)	4 (9.3)
p.Gly13Asp (G13D)	2 (7.4)	4 (9.3)
MSI status
MSS + MSI-L	25 + 2 (100)	34 + 2 (83.7)	0.0382a
MSI-H	0	7 (16.3)

Fisher's exact test.

^aP value < 0.05 were considered statistically significant. WT: Wild; MSI: Microsatellite instability; MSS: Microsatellite stable; MSI-L: Microsatellite instability low; MSI-H: Microsatellite instability high; CRA: Colorectal adenoma; CRC: Colorectal cancer.