Pathological significance of abnormal recepteur d’origine nantais and programmed death ligand 1 expression in colorectal cancer

Table 1 Comparison of RON and PD-L1 expression and clinicopathological characteristics of patients with colorectal cancer in the FAHZUSM and Gene Expression Omnibus cohorts, n (%)

	FAHZUSM cohort (n = 381)										GEO cohort (n = 287)
Characteristic	Cases	RON expression			PD-L1 expression						Cases No (%)	RON expression			PD-L1 expression
		Low	High	1P value	Low	High (TIMC)	2P value	Low	High (TC)	3P value		Low	High	1P value	Low	High (TC)	3P value
No. of cases number	381	260	121		247	91		247	43		287	193	94		48	239
Age (mean ± SD)	61.16 ± 12.419	60.40 ± 12.560	62.81 ± 11.997	0.859	60.94 ± 12.37	62.04 ± 13.080	0.771	60.94 ± 12.37	61.16 ± 12.419	0.938	68.44 ± 14.00	66.87 ± 13.18	71.65 ± 15.12	0.006	66.94 ± 14.33	68.74 ± 13.95	0.417
Sex
Man	215 (56.4)	157 (60.4)	58 (47.9)	0.023	143 (57.9)	54 (59.3)	0.811	143 (57.9)	18 (41.9)	0.051	158 (55.1)	111 (57.5)	47 (50)	0.230	30 (62.5)	128 (53.6)	0.256
Women	166 (43.6)	103 (39.6)	63 (52.1)		104 (42.1)	37 (40.7)		104 (42.1)	25 (58.1)		129 (44.9)	82 (42.5)	47 (50)		18 (37.5)	111 (46.4)
Principal diagnosis
Rectum	229 (60.1)	158 (61.1)	71 (58.7)	0.698	153 (61.9)	47 (51.6)	0.088	153 (61.9)	29 (67.4)	0.491	165 (57.5)	111 (57.5)	54 (57.4)	0.920	34 (70.8)	131 (54.8)	0.040
Colon	152 (39.9)	102 (38.9)	50 (41.3)		94 (38.1)	44 (48.4)		94 (38.1)	14 (32.6)		122 (42.5)	82 (42.5)	40 (42.6)		14 (29.2)	108 (45.2)
Pathological grade
Well/moderate	9 (2.4)	6 (2.3)	3 (2.5)	0.177	5 (2)	3 (3.3)	0.778	5 (2)	1 (2.3)	0.898	NA
Poor	346 (90.8)	232 (89.2)	114 (94.2)		224 (90.7)	82 (90.1)		224 (90.7)	40 (93)
Unknown	26 (6.8)	22 (8.5)	4 (3.3)		18 (7.1)	6 (6.3)		18 (7.3)	2 (4.7)
T stage
Tis-T2	107 (28.1)	85 (32.7)	22 (18.2)	0.001	73 (29.6)	23 (25.3)	0.105	73 (29.6)	11 (25.6)	0.864	28 (9.7)	18 (9.3)	10 (10.6)	0.459	3 (6.3)	25 (10.5)	0.437
T3	177 (46.5)	121 (46.5)	56 (46.3)		118 (47.8) (46.8%)	37 (40.7)		118 (47.8)	22 (51.2)		208 (72.5)	144 (74.6)	64 (68.1)20		39 (81.3)	169 (70.7)
T4	97 (25.5)	54 (20.8)	43 (35.5)		56 (22.7)	31 (34.1)		56 (22.7)	10 (23.3)		51 (17.8)	31 (16.1)	20 (21.3)		6 (12.5)	45 (18.8)
N stage
N0	251 (65.9)	184 (70.8)	67 (55.4)	0.003	164 (66.4)	62 (68.1)	0.764	164 (66.4)	25 (58.1)	0.294	141 (49.1)	97 (50.3)	44 (46.8)	0.583	27 (56.3)	114 (44.7)	0.279
N (1-2)	91 (23.9)	76 (29.2)	54 (44.6)		83 (33.6)	29 (31.9)		83 (33.6)	12 (27.9)		146 (50.9)	96 (49.7)	50 (53.2)		21 (43.7)	125 (52.3)
M stage
M0	368 (96.6)	252 (96.9)	116 (95.9) (96.3%)	0.822	241 (97.6)	85 (93.4)	0.133	241 (97.6)	42 (97.7)	1.000	268 (93.4)	183 (94.8)	85 (90.4)	0.160	47 (97.9)	221 (92.5)	0.216
M1	13 (3.4)	8 (3.1)	5 (4.1)		6 (2.4)	6 (6.6)		6 (2.4)	1 (2.3)		19 (6.6)	10 (5.2)	9 (9.6)		1 (2.1)	18 (7.5)
Disease stage
0-II	246 (64.6)	180 (69.2)	66 (54.5)	0.016	162 (65.6)	59 (64.8)	0.897	162 (65.6)	25 (58.1)	0.346	139 (48.4)	95 (49.2)	44 (46.8)	0.701	27 (56.3)	112 (46.9)	0.235
III-IV	135 (35.4)	80 (30.8)	55 (45.5)		85 (34.4)	32 (35.2)		85 (34.4)	18 (41.9)		148 (51.6)	98 (50.8)	50 (53.2)		21 (43.8)	127 (53.1)
Histological type
Adenocarcinoma	335 (87.9)	227 (87.3)	108 (89.3)	0.706	214 (86.6)	84 (92.3)	0.130	214 (86.6)	37 (86)	0.488	287 (100)	193 (100)	94 (100)		48 (100)	239 (100)
Mucinous/SRCC	36 (9.4)	25 (9.6)	11 (9.1)		28 (11.3)	4 (4.4)		28 (11.3)	4 (9.3)		0 (0)	0 (0)	0 (0)		0 (0)	0 (0)
Other	10 (2.6)	8 (3.1)	2 (1.7)		5 (2.0)	3 (3.3)		5 (2.0)	2 (4.7)		0 (0)	0 (0)	0 (0)		0 (0)	0 (0)
Treatment
Yes	173 (45.4)	110 (42.3)	63 (52.1)	0.043	112 (45.3)	38 (41.8)	0.694	112 (45.3)	23 (32.6)	0.472	115 (40.1)	89 (46.1)	26 (27.7)	0.003	22 (45.8)	93 (38.9)	0.372
No	158 (41.5)	119 (45.8)	39 (32.2)		105 (42.5)	39 (42.9)		105 (42.5)	14 (53.5)		172 (59.9)	104 (53.9)	68 (72.3)		26 (54.2)	146 (61.1)
Unknown	50 (13.1)	31 (11.9)	19 (15.7)		30 (12.1)	14 (15.4)		30 (12.1)	6 (14)			0 (0)	0 (0)
PD-L1 (protein)
Low	247 (64.8)	177 (73.1)	70 (26.9)	0.9764							NA
High (TIMCs)	91 (23.9)	65 (26.9)	26 (27.1)
High (TCs)	43 (11.3)	18 (9.2)	25 (26.3)	< 0.0015
PD-L1 (mRNA)
Low	NA										48 (16.7)	31 (16.1)	17 (18.1)	0.666
High											239 (83.3)	162 (83.9)	77 (81.9)

¹Comparing RON high expression and RON low expression.

²Comparing PD-L1 low expression and PD-L1 expression in TIMCs.

³Comparing PD-L1 low expression and PD-L1 expression in TCs.

⁴PD-L1 low (both TCs and TIMCs) vs PD-L1 high (TIMCs).

⁵PD-L1 low (both TCs and TIMCs) vs PD-L1 high TCs. Bold entries indicate statistical significance (P < 0.05). GEO: The Gene Expression Omnibus; RON: Recepteur d’origine nantais; PD-L1: Programmed death ligand 1; TCs: Tumor cells; TIMCs: Tumor-infiltrating mononuclear cells; FAHZUSM: The First Affiliated Hospital, Zhejiang University School of Medicine.

Table 2 Comparison of RON (tumor cells) and PD-L1 (tumor cells) expression and clinicopathological characteristics of patients with colorectal cancer in the FAHZUSM cohort, n (%)

Characteristic	Cases	RON and PD-L1 (TCs) expression in the tumor microenvironment
		RON low/PD-L1 low	RON high/PD-L1 low	RON low/PD-L1 high	RON high/PD-L1 high	P value
Number of cases	290	177	70	18	25
Age (mean ± SD)	60.89 ± 12.215	60.37 ± 12.299	62.40 ± 12.518	58.39 ± 12.930	62.12 ± 10.179	0.495
Sex
Man	161 (55.5)	108 (61)	35 (50.0)	9 (50.0)	9 (36.0)	0.061
Women	129 (44.5)	68 (39)	35 (50.0)	9 (50.0)	16 (64.0)
Principal diagnosis				s
Rectum	182 (62.8)	116 (65.5)	37 (52.9)	14 (77.8)	15 (60.0)	0.149
Colon	108 (37.2)	61 (34.5)	33 (47.1)	4 (22.2)	10 (40.0)
Pathological grade
Well/moderate	6 (2.1)	3 (1.7)	2 (2.9)	0 (0.0)	1 (4.0)	0.264
Poor	264 (9.1)	158 (89.3)	66 (94.3)	16 (88.9)	24 (96.0)
Unknown	20 (6.9)	16 (9.0)	2 (2.9)	2 (11.1)	0 (0.0)
T stage
Tis-T2	84 (29)	64 (36.2)	9 (12.9)	5 (27.8)	6 (24.0)	0.003
T3	140 (48.3)	82 (46.3)	36 (51.4)	11 (61.1)	11 (44.0)
T4	66 (22.8)	31 (17.5)	25 (35.7)	2 (11.1)	8 (32.0)
N stage
N0	238 (66.9)	125 (70.6)	39 (55.7)	12 (66.7)	13 (66.4)	0.072
N (1-2)	118 (33.1)	52 (29.4)	31 (44.3)	6 (33.3)	12 (33.6)
M stage
M0	283 (97.6)	173 (97.7)	68 (97.1)	18 (100)	24 (96.0)	0.735
M1	7 (2.4)	4 (2.3)	2 (2.9)	0 (0.0)	1 (4.0)
Stage
0-II	187 (64.5)	123 (69.5)	39 (55.7)	12 (66.7)	13 (52.0)	0.110
III-IV	103 (35.5)	54 (30.5)	31 (33.3)	6 (33.3)	12 (48.0)
Histological type
Adenocarcinoma	251 (86.6)	152 (85.9)	62 (88.6)	15 (83.3)	22 (88.0)	0.493
Mucinous/SRCC	32 (11)	21 (11.9)	7 (10.0)	1 (5.6)	3 (12.0)
Other	7 (2.4)	4 (2.3)	1 (1.4)	2 (11.1)	0 (0.0)
Treatment
Yes	135 (46.6)	83 (46.9)	22 (31.4)	6 (33.3)	12 (48.0)	0.158
No	119 (41.0)	74 (41.8)	38 (54.3)	11 (61.1)	8 (32.0)
Unknown	36 (12.4)	20 (11.3)	10 (14.3)	1 (5.6)	5 (20.0)

Bold entries indicate statistical significance (P < 0.05). RON: Recepteur d’origine nantais; PD-L1: Programmed death ligand 1; TCs: Tumor cells; FAHZUSM: The First Affiliated Hospital, Zhejiang University School of Medicine.

Table 3 Comparison of RON (tumor cells) and PD-L1 (tumor-infiltrating mononuclear cells) expression and clinicopathological characteristics of patients with colorectal cancer in the FAHZUSM cohort, n (%)

Characteristic	Cases	RON and PD-L1 (TIMCs) expression in the tumor microenvironment
		RON low/PD-L1 low	RON high/PD-L1 low	RON low/PD-L1 high	RON high/PD-L1 high	P value
Number of cases	338	177	70	65	26
Age (mean ± SD)	61.24 ± 12.555	60.37 ± 12.299	62.40 ± 12.518	61.03 ± 13.286	64.58 ± 12.436	0.348
Sex
Man	197 (58.3)	108 (61)	35 (50.0)	40 (61.5)	14 (52.8)	0.368
Women	141 (41.7)	68 (39)	35 (50.0)	25 (38.5)	12 (46.2)
Principal diagnosis
Rectum	200 (59.2)	116 (65.5)	37 (52.9)	28 (43.1)	19 (73.1)	0.004
Colon	138 (40.8)	61 (34.5)	33 (47.1)	37 (56.9)	7 (26.9)
Pathological grade
Well/moderate	8 (2.4)	3 (1.7)	2 (2.9)	3 (4.6)	0 (0)	0.492
Poor	306 (90.5)	158 (89.3)	66 (94.3)	58 (89.2)	24 (92.3)
Unknown	24 (7.1)	16 (9.0)	2 (2.9)	4 (6.2)	2 (7.7)
T stage
Tis-T2	96 (28.4)	64 (36.2)	9 (12.9)	16 (2.6)	7 (26.9)	0.002
T3	155 (45.9)	82 (46.3)	36 (51.4)	28 (43.1)	9 (34.6)
T4	87 (25.7)	31 (17.5)	25 (35.7)	21 (32.3)	10 (38.5)
N stage
N0	226 (66.9)	125 (70.6)	39 (55.7)	47 (72.3)	15 (57.7)	0.075
N (1-2)	112 (33.1)	52 (29.4)	31 (44.3)	18 (27.7)	11 (42.3)
M stage
M0	326 (96.4)	173 (97.7)	68 (97.1)	61 (93.8)	24 (92.3)	0.235
M1	12 (3.6)	4 (2.3)	2 (2.9)	4 (2.3)	2 (7.7)
Stage
0-II	221 (65.4)	123 (69.5)	39 (55.7)	45 (69.2)	14 (53.8)	0.104
III-IV	117 (34.6)	54 (30.5)	31 (33.3)	20 (30.8)	12 (46.2)
Histological type
Adenocarcinoma	287 (84.9)	152 (85.9)	62 (88.6)	60 (92.3)	13 (50)	< 0.001
Mucinous/SRCC	40 (11.8)	21 (11.9)	7 (10.0)	3 (4.6)	9 (34.6)
Other	11 (3.3)	4 (2.3)	1 (1.4)	2 (3.1)	4 (15.4)
Treatment
Yes	298 (88.2)	83 (46.9)	22 (31.4)	30 (46.2)	24 (92.3)	< 0.001
No	32 (9.5)	74 (41.8)	38 (54.3)	25 (38.5)	1 (3.8)
Unknown	8 (2.4)	20 (11.3)	10 (14.3)	10 (15.4)	1 (3.8)

Bold entries indicate statistical significance (P < 0.05). RON: Recepteur d’origine nantais; PD-L1: Programmed death ligand 1; TCs: Tumor cells; TIMCs: Tumor-infiltrating mononuclear cells; FAHZUSM: The First Affiliated Hospital, Zhejiang University School of Medicine.

Table 4 Univariate and multivariate Cox proportional hazard analyses of overall survival in patients with colorectal cancer in the FAHZUSM and Gene Expression Omnibus cohorts

Variable	FAHZUSM cohort				GEO cohort
	Univariate analysis, HR (95%CI)	P value	Multivariate analysis, HR (95%CI)	P value	Univariate analysis, HR (95%CI)	P value	Multivariate analysis, HR (95%CI)	P value
Age (mean ± SD)	1.029 (1.011-1.048)	0.002	1.031 (1.011-1.050)	0.002	1.025 (1.009-1.041)	0.002	1.028 (1.012-1.045)	0.001
Gender
Male	1.000	0.011	1.000	0.008	1.000	0.293	1.000	0.076
Female	0.562 (0.362-0.874)		0.527 (0.328-0.847)		0.814 (0.555-1.194)		0.698 (0.469-1.039)
Principal diagnosis
Rectum	1.000	0.298	1.000	0.318	1.000	0.688	1.000	0.544
Colon	0.796 (0.518-1.224)		0.790 (0.497-1.255)		1.081 (0.738-1.585)		0.882 (0.589-1.322)
Histological type
Adenocarcinoma	1.000	0.408	1.000	0.387	NA
Mucinous/SRCC	2.464 (0.343-17.710)		2.720 (0.263-28.122)
Unknown	3.355 (0.433-25.986)		1.232 (0.129-11.795)
T stage
Tis-T2	1.000	< 0.001	1.000	0.003	1.000	0.001	1.000	0.007
T3	0.211 (0.108-0.413)		0.298 (0.141-0.632)		0.333 (0.145-0.766)		0.381 (0.159-0.914)
T4	0.503 (0.325-0.781)		0.596 (0.361-0.897)		0.455 (0.292-0.709)		0.476 (0.295-0.769)
N stage
N0	1.000	< 0.001	1.000	0.111	1.000	0.062	1.000	0.141
N (1-2)	3.156 (2.083-4.783)		5.376 (0.678-42.660)		1.435 (0.982-2.097)		0.306 (0.063-1.479)
M stage
M0	1.000	< 0.001	1.000	0.039	1.000	< 0.001	1.000	0.008
M1	4.338 (2.094-8.985)		2.445 (1.048-5.701)		3.546 (2.013-6.247)		2.481 (1.269-4.852)
Stage
0-II	1.000	< 0.001	1.000	0.406	1.000	0.029	1.000	0.099
III-IV	3.102 (2.044-4.708)		0.409 (0.050-3.366)		1.530 (1.045-2.242)		3.924 (0.774-19.888)
Pathological grade
Well/moderate	1.000	< 0.001	1.000	0.003	NA
Poor	7.689 (2.434-24.290)		4.044 (1.103-14.830)
Unknown	1.266 (0.513-3.126)		0.375 (0.104-1.357)
RON (protein)
Low	1.000	0.001	1.000	0.012	1.000	0.035	1.000	0.466
High	2.060 (1.364-3.112)		1.788 (1.138-2.808)		1.515 (1.030-2.228)		1.170 (0.767-1.785)
RON (mRNA)
Low	NA				1.000	0.035	1.000	0.466
High					1.515 (1.030-2.228)		1.170 (0.767-1.785)
PD-L1 (protein)
Low	1.000	< 0.001	1.000	0.001	NA
High (TIMCs)	0.544 (0.293-1.011)		0.676 (0.350-1.304)
High (TCs)	1.325 (0.669-2.511)		1.654 (0.819-3.338)
PD-L1 (mRNA)
Low	NA				1.000	0.047	1.000	0.083
High					1.835 (1.007-3.345)		1.719 (0.931-3.173)

All variables are adjusted by the Cox proportional hazard model. Bold entries indicate statistical significance (P < 0.05). GEO: The Gene Expression Omnibus; RON: Recepteur d’origine nantais; PD-L1: Programmed death ligand 1; TCs: Tumor cells; TIMCs: Tumor-infiltrating mononuclear cells; FAHZUSM: The First Affiliated Hospital, Zhejiang University School of Medicine.