Large tumor suppressor 2 is a prognostic biomarker and correlated with immune infiltrates in colorectal cancer

Figures & data

Figure 1. LATS2 mRNA expression in CRC tissues and cell lines. (a) LATS2 mRNA expression was significantly decreased in CRC tissues compared with matched adjacent normal tissues. (b-c) In the TCGA database, LATS2 mRNA expression was markedly lower in COAD and READ tissues compared with normal tissues. (d) LATS2 expression was decreased in CRC cell lines compared with a normal intestinal epithelial cell (NCM460). Data were expressed as mean ± SD, *P < 0.05; **P < 0.01; ***P < 0.001

Table 1. LATS2 expression in colorectal benign and malignant tissues

Characteristic	n	LATS2 High expression (%)	Low or no expression (%)	Pearson χ^2	P value
Chronic colitis	26	6 (23.08)	20 (76.92)
Low-grade intraepithelial neoplasia	44	7 (15.91)	37 (84.09)
High-grade intraepithelial neoplasia	22	2 (9.09)	20 (90.91)
Cancer	213	72 (33.80)	141 (66.20)
Surgical margin ^a	174	98 (56.32)	76 (43.68)
Total	479	185 (38.62)	294 (61.38)	45.401	<0.001***

^aEpithelium without intraepithelial neoplasia from colorectal cancer. ***P < 0.001.

Figure 2. Representative patterns of LATS2 protein expression in colorectal benign and malignant tissues on TMA sections. (a–b) Colorectal cancer with low or no LATS2 expression. (c–d) Colorectal cancer with high LATS2 expression. (e–f) High-grade intraepithelial neoplasia with low LATS2 expression. (g–h) Low-grade intraepithelial neoplasia with low LATS2 expression. (i–j) Normal surgical margin of colorectal cancer with high LATS2 expression

Table 2. Associations of LATS2 protein expression with clinicopathological characteristics in colorectal cancer patients

Characteristic	n	LATS2 High expression (%) Low or no expression (%)		Pearson χ^2	P value
Total	213	72 (33.80)	141 (66.20)
Sex				0.627	0.428
Male	135	43 (31.85)	92 (68.15)
Female	78	29 (37.18)	49 (62.82)
Age				0.041	0.839
<60	72	25 (34.72)	47 (65.28)
≥60	141	47 (33.33)	94 (66.67)
Location				2.712	0.100
Colon	160	59 (36.88)	101 (63.12)
Rectum	53	13(24.53)	40 (75.47)
Differentiation				12.555	0.002**
Poor	22	0 (0)	22 (100)
Well and middle	177	67 (37.85)	110 (62.15)
Others ^a	14	5 (35.71)	9 (64.29)
TNM stage				12.216	0.002**
0-I	46	23 (50.00)	23 (50.00)
II	85	32 (37.65)	53 (62.35)
III+IV	82	17 (20.73)	65 (79.27)
T				4.156	0.041*
Tis+T1+ T2	53	24 (45.28)	29 (54.72)
T3,4b	160	48 (30.00)	112 (70.00)
N				10.972	0.012*
N0	134	53 (39.55)	81 (60.45)
N1a	39	9 (23.08)	30 (76.92)
N1b	21	7 (33.33)	14 (66.67)
N2a,b	19	1 (5.26)	18 (94.74)
M				7.652	0.006**
M0	199	72 (36.18)	127 (63.82)
M1a+1b	14	0(0)	14 (100)
Preoperative CEA (ng/ml)				1.263	0.532
≤5	131	48 (36.64)	83 (63.36)
>5	26	8 (30.77)	18 (69.23)
unknown	56	16 (28.57)	40(71.43)

^aMucinous adenocarcinoma, 14 cases. *P < 0.05; **P < 0.01

Table 3. Univariate and multivariate analyses of prognostic factors for overall survival in colorectal cancer patients

	Univariate analysis			Multivariate analysis
	HR	p>|z|	95%CI	HR	p>|z|	95%CI
LATS2 expression High vs. Low	0.087	<0.001***	0.035–0.216	0.151	<0.001***	0.053–0.429
Age (years) ≤60 vs. >60	0.954	0.840	0.602–1.510
Sex Male vs. Female	1.293	0.283	0.809–2.067
Location Colon vs. Rectum	1.388	0.174	0.866–2.227
Differentiation Poor vs. Well and middle	2.672	<0.001***	1.630–4.379	1.411	0.251	0.783–2.543
TNM stage 0-I vs. II vs. III+IV	2.193	<0.001***	1.575–3.054	1.922	0.048*	1.377–2.993
T Tis+T1+ T2 vs. T3, 4b	6.693	<0.001***	2.705–16.560
N N0 vs. N1a vs. N1b vs. N2a, b	1.479	<0.001***	1.213–1.804
M M0 vs. M1a+1b	6.418	<0.001***	3.470–11.869
Preoperative CEA, ng/ml ≤5 vs. >5	2.593	0.001**	1.501–4.478	2.089	0.052	1.066–4.093

Abbreviation: HR, hazard ratio; CI, confidence interval. *P < 0.05; **P < 0.01; ***P < 0.001.

Figure 3. Survival analysis of CRC patients by the Kaplan–Meier method. (a) Overall survival in patients with high LATS2 expression was significantly higher than that in patients with low LATS2 expression. (b) Overall survival in patients with stage II or stage III–IV CRC was significantly lower than that in patients with stage 0–I CRC

Figure 4. Notable immune-related signaling pathways in the high LATS2 expression group. (a) LATS2-activated signaling pathways. (b) Chemokine signaling pathways. (c) Cytokine-cytokine receptor interaction signaling pathways. (d) JAK-STAT signaling pathways. (e) Intestinal immune network for IgA production signaling pathways

Abbreviation: FDR, false discovery rate; NES, normalized enrichment score.

Table 4. Correlations between LATS2 expression and marker genes for tumor-infiltrating immune cells using the TIMER database

		COAD				READ
Description	Gene markers	None		Purity		None		Purity
		Cor	P	Cor	P	Cor	P	Cor	P
CD8 + T cell	CD8A	0.32	***	0.23	***	0.24	***	0.07	ns
	CD8B PTPRC	0.21 0.54	*** ***	0.16 0.47	** ***	0.18 0.47	* ***	0.05 0.34	ns ***
T cell (general)	CD3D	0.25	***	0.13	**	0.11	ns	−0.06	ns
	CD3E	0.36	***	0.26	***	0.26	***	0.09	ns
	CD2	0.34	***	0.25	***	0.25	**	0.10	ns
B cell	CD19	0.22	***	0.12	*	0.11	ns	0.01	ns
	CD79A	0.31	***	0.20	***	0.22	**	0.04	ns
Monocyte	CD14	0.53	***	0.46	***	0.44	***	0.34	***
	CD115 (CSF1R)	0.57	***	0.50	***	0.54	***	0.44	***
TAM	CD68	0.48	***	0.43	***	0.45	***	0.37	***
	IL10	0.42	***	0.36	***	0.32	***	0.19	*
M1 Macrophage	INOS (NOS2)	−0.14	**	−0.17	***	−0.14	ns	−0.13	ns
	CD80	0.42	***	0.36	***	0.38	***	0.26	**
	IRF5	0.35	***	0.35	***	0.34	***	0.33	***
	IL6	0.39	***	0.33	***	0.40	***	0.36	***
	CD64 (FCGR1A)	0.41	***	0.33	***	0.35	***	0.28	***
M2 Macrophage	CD206	0.48	***	0.41	***	0.46	***	0.33	***
	VSIG4	0.51	***	0.43	***	0.44	***	0.36	***
	MS4A4A	0.52	***	0.45	***	0.52	***	0.45	***
Neutrophils	CD66b	−0.18	***	−0.17	***	−0.21	**	−0.09	ns
	CCR7	0.40	***	0.32	***	0.26	***	0.16	ns
Natural killer cell	KIR2DL1	0.11	*	0.04	ns	0.15	ns	0.10	ns
	KIR2DL3	0.12	*	0.08	ns	0.08	ns	0.01	ns
Dendritic cell Th1 Th2 Tfh Th17 Treg T cell exhaustion	KIR2DL4 KIR3DL1 KIR3DL2 BDCA-1 (CD1C) BDCA-3 (THBD) BDCA-4 (NRP1) IL3RA STAT4 STAT1 T-bet (TBX21) GATA3 STAT6 IL13 IL21 BCL6 STAT3 IL17A FOXP3 CD25 PD-1 (PDCD1) CTLA4	0.18 0.18 0.22 0.37 0.67 0.71 0.52 0.38 0.44 0.36 0.41 0.23 0.29 0.18 0.62 0.46 –0.18 0.46 0.45 0.31 0.39	*** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** ***	0.11 0.11 0.15 0.29 0.64 0.67 0.48 0.29 0.39 0.29 0.36 0.24 0.24 0.13 0.59 0.43 –0.19 0.39 0.36 0.23 0.32	* * ** *** *** *** *** *** *** *** *** *** *** ** *** *** **** *** *** *** ***	0.15 0.05 0.18 0.26 0.70 0.70 0.48 0.30 0.44 0.32 0.45 0.20 0.12 –0.03 0.54 0.42 –0.27 0.49 0.45 0.27 0.39	ns ns * *** *** *** *** *** *** *** *** * ns ns *** *** *** *** *** *** ***	−0.03 –0.04 0.06 0.13 0.64 0.67 0.45 0.17 0.31 0.18 0.38 0.23 0.01 –0.09 0.54 0.39 –0.25 0.39 0.35 0.10 0.23	ns ns ns ns *** *** *** * *** * *** ** ns ns *** *** ** *** *** ns **
	LAG3	0.34	***	0.25	****	0.24	**	0.10	ns

*P < 0.05; **P < 0.01; ***P < 0.001.

Figure 5. Correlations of LATS2 expression with tumor microenvironment and immune infiltration levels in CRC. (a) Positive correlation between LATS2 expression and immune score in CRC. (b) LATS2 was positively correlated with tumor-infiltrating immune cells in COAD by TIMER. (c) LATS2 was positively correlated with tumor-infiltrating immune cells in READ by TIMER. (d) LATS2 was positively correlated with tumor-infiltrating immune cells in COAD by ssGSEA. (e) LATS2 was positively correlated with tumor-infiltrating immune cells in READ by ssGSEA

Figure 6. Correlations of LATS2 expression with NRP1, BCL6, PTPRC, and THBD expression in cancer samples and analysis of the PPI network for LATS2. (a–d) LATS2 was positively associated with NRP1, BCL6, PTPRC, and THBD expression in almost all cancers examined, based on data from the TCGA. (e) Protein-protein network view in the GeneMANIA dataset showing the interaction networks of LATS2

Data availability statement

The gene expression data of CRC and normal tissues were extracted from the TCGA Data Portal (https://tcga-data.nci.nih.gov/tcga). All the data will be provided on reasonable request from the corresponding author.