The Perspective of Diagnostic and Prognostic Values of Lipoxygenases mRNA Expression in Colon Adenocarcinoma

Figures & data

Table 1 Baseline Patient Characteristics in TCGA Cohort

Variables	RFS					OS
	Patients (n=371)	No. of Events	MST (days)	HR (95%CI)	Log-rank P	Patients (n=438)	No. of Events	MST (days)	HR (95%CI)	Log-rank P
Age (years)
<65	145	31		1	0.402	166	39	3042	1	0.371
≥65	224	46		1.216 (0.769–1.923)		270	59	1910	1.204 (0.801–1.810)
Missing^a	2					2
Gender
Male	201	48	2270	1	0.046	234	54	2475	1	0.545
Female	170	30	NA	0.629 (0.398–0.996)		204	44	NA	0.884 (0.593–1.318)
TNM Stage
I	61	5	NA	1	<0.001	73	4	NA	1	<0.001
II	144	28	NA	1.934 (0.745–5.020)		168	28	2821	2.308 (0.807–6.602)
III	109	25	NA	2.829 (1.082–7.394)		126	31	NA	1.446 (1.446–11.634)
IV	47	18	1069	6.614 (2.452–17.840)		61	31	858	11.355 (4.003–32.208)
Missing^b	10					10
CEA
≥5 ng/mL	81	24	1678	1	0.005	98	30	1711	1	<0.001
<5 ng/mL	154	23	NA	0.452 (0.255–0.802)		180	21	3042	0.326 (0.185–0.573)
Missing^c	136					260
Tumor
location
Right	212	41	NA	1	0.820	251	62	2532	1	0.069
Left	143	32	2270	1.055 (0.664–1.676)		171	30	3042	0.669 (0.433–1.035)
Missing^d	16					16
KRAS
Yes	44	19	1069	1	0.005	48	25	1518	1	0.001
No	293	53	2564	0.474 (0.277–0.809)		355	58	2821	0.455 (0.280–0.737)
Missing^e	34					35
Lymphatic invasion
Yes	212	34	NA	1	<0.001	243	42	NA	1	<0.001
No	128	38	1801	2.322 (1.452–3.713)		152	44	2003	0.468 (0.304–0.718)
Missing^f	31					43

Notes: ^aInformation of age was unknown for both RFS and OS in two patients. ^bInformation of TNM stage was unknown for both RFS and OS in 10 patients. ^cInformation of CEA was unavailable in 136 patients of RFS and 260 patients of OS. ^dInformation of tumor location was unavailable for both RFS and OS in two patients. ^eInformation of KRAS was unavailable in 34 patients of RFS and 35 patients of OS. ^fInformation of lymphatic invasion was not reported in 31 patients of RFS and 43 patients of OS.

Abbreviations: RFS, recurrence-free survival; OS, overall survival; TCG, The Cancer Genome Atlas; MST, median survival time; NA, not available; CEA, carcino-embryonic antigen.

Figure 1 Pearson’s correlations analysis and gene-gene and protein-protein interaction networks of ALOX genes. (A) Pearson correlation analysis of ALOX genes. Red indicates positive correlation and blue indicates a negative correlation. (B) GeneMANIA interaction networks; (C) Protein‑protein interaction networks.

Abbreviation: ALOX, lipoxygenases.

Figure 2 Gene expression distribution of ALOX genes in TCGA. *P<0.05; **P<0.01; ***P<0.0001.

Abbreviations: NS, not significant; ALOX, lipoxygenases; TCGA, The Cancer Genome Atlas.

Figure 3 Expression levels of ALOX genes in tumor and normal tissues from the GEPIA website. (A), ALOXE3; (B), ALOX5; (C), ALOX12; (D), ALOX12B; (E), ALOX15; (F), ALOX15B.

Abbreviations: ALOX, lipoxygenases; COAD, colon adenocarcinoma; GEPIA, Gene Expression Profiling Interactive Analysis.

Figure 4 Expression levels of ALOX genes in tumor and normal tissues from the MERAV website. (A), ALOXE3; (B), ALOX5; (C), ALOX12; (D), ALOX12B; (E), ALOX15; (F), ALOX15B.

Abbreviations: ALOX, lipoxygenases; MERAV, metabolic gene rapid visualizer.

Figure 5 The diagnostic ROC curves of ALOX genes distinguish COAD tumor tissues and adjacent normal tissues in the TCGA cohort. ROC curves of ALOXE3 (A); ALOX5 (B); ALOX12 (C); ALOX12B (D); ALOX15 (E); ALOX15B (F).

Abbreviations: ALOX, lipoxygenases; COAD, colon adenocarcinoma; TCGA, The Cancer Genome Atlas; ROC, receiver operating characteristic; AUC, area under the curve.

Figure 6 The verification of the relative expression of ALOXE3 and ALOX12 in COAD. (A) The relative expression of ALOXE3 between tumor tissues and adjacent normal tissues. (B) The diagnostic ROC curve of ALOXE3 in COAD. (C) The relative expression of ALOX12 between tumor tissues and adjacent normal tissues. (D) The diagnostic ROC curve of ALOX12 in COAD.

Abbreviations: ALOX, lipoxygenases; COAD, colon adenocarcinoma; TCGA, The Cancer Genome Atlas.

Table 2 Prognostic Values of ALOX Gene Expressions in COAD of TCGA Cohort

Gene Expression	RFS							OS
	Patients (n=371)	No. of Events	MST (days)	Crude HR (95%CI)	Crude P	Adjusted HR (95% CI)	Adjusted P^a	Patients (n=438)	No. of Events	MST (days)	Crude HR (95%CI)	Crude P	Adjusted HR (95%CI)	Adjusted P^b
ALOXE3
Low	183	33	NA	1	0.142	1	0.382	219	38	2475	1	0.029	1	0.025
High	188	45	2394	1.398 (0.892–2.192)		1.272 (0.741–2.184)		219	60	2532	1.569 (1.044–2.358)		1.765 (1.074–2.901)
ALOX5
Low	183	36	NA	1	0.557	1	0.368	219	48	NA	1	0.751	1	0.085
High	188	42	2564	1.143 (0.732–1.785)		1.293 (0.739–2.262)		219	50	2532	1.066 (0.717–1.586)		1.552 (0.942–2.556)
ALOX12
Low	189	32	NA	1	0.016	1	0.753	219	35	NA	1	0.001	1	0.046
High	182	46	2270	0.916 (0.584–1.436)		1.731 (1.102–2.719)		219	63	1849	2.015 (1.333–3047)		1.680 (1.009–2.796)
ALOX12B
Low	195	36	NA	1	0.121	1	0.604	219	45	NA	1	0.129	1	0.070
High	176	42	2176	1.420 (0.909–2.219)		1.150 (0.678–1.953)		219	53	2003	1.359 (0.913–2.024)		1.553 (0.965–2.499)
ALOX15
Low	191	38	NA	1	0.311	1	0.722	219	47	2821	1	0.311	1	0.924
High	180	40	2564	1.260 (0.805–1.971)		1.104 (0.640–1.905)		219	51	2003	1.229 (0.824–1.833)		1.024 (0.631–1.662)
ALOX15B
Low	192	42	NA	1	0.978	1	0.423	219	50	2532	1	0.773	1	0.828
High	179	36	2564	1.066 (0.644–1.572)		0.794 (0.451–1.397)		219	48	2034	1.060 (0.712–1.578)		1.055 (0.651–1.708)

Notes: ^aAdjustment for gender, TNM stage, CEA, KRAS and lymphatic invasion. ^bAdjustment for TNM stage, CEA, KRAS and lymphatic invasion.

Abbreviations: COAD, colon adenocarcinoma; ALOX, lipoxygenases; TCGA, The Cancer Genome Atlas; OS, overall survival; RFS, recurrence-free survival; MST, median survival time; NA, not available; CEA, carcino-embryonic antigen.

Figure 7 Kaplan–Meier survival curves for ALOX genes in COAD of the TCGA cohort. OS stratified by ALOXE3 (A); ALOX5 (B); ALOX12 (C); ALOX12B (D); ALOX15 (E); ALOX15B (F).

Abbreviations: ALOX, lipoxygenases; COAD, colon adenocarcinoma; TCGA, The Cancer Genome Atlas; OS, overall survival.

Table 3 Joint Effects Analysis of ALOXE3 and ALOX12 Expressions in COAD Patients OS

Group	ALOXE3	ALOX12	Patients (n=438)	No. of Event	MST (days)	Crude HR (95%CI)	Crude P	Adjusted HR (95%CI)	Adjusted P^a
A	Low	Low	120	17	NA	1	0.003	1	0.027
B	Low	High	99	21	1849	1.873 (0.987–3.552)	0.055	1.263 (0.573–2.780)	0.563
C	High	Low	99	18	NA	1.396 (0.719–2.709)	0.325	1.324 (0.600–2.918)	0.487
D	High	High	120	42	1711	2.710 (1.542–4.765)	0.01	2.472 (1.248–4.900)	0.009

Notes: ^aAdjusted for gender, TNM stage, CEA, KRAS and lymphatic invasion.

Abbreviations: ALOX, lipoxygenases; OS, overall survival; MST, median survival time; TCGA, The Cancer Genome Atlas; NA, not available. CEA, carcino-embryonic antigen.

Figure 8 Joint effects analysis of ALOXE3 and ALOX12 of COAD OS in the TCGA cohort.

Abbreviations: ALOX, lipoxygenases; COAD, colon adenocarcinoma; TCGA, The Cancer Genome Atlas; OS, overall survival.

Figure 9 Risk score model and ROC curves. (A) Risk score model plot including risk score ranking, survival status, and heatmap. (B) Kaplan–Meier curves for low- and high-risk groups. (C) ROC curves for one‑, two‑, three‑, four‑ and five‑year survival rates from the risk score model.

Abbreviations: ALOX, lipoxygenases; ROC, receiver operating characteristic; COAD, colon adenocarcinoma.

Figure 10 Nomogram for predicting the prognosis of COAD patient.

Abbreviations: COAD, colon adenocarcinoma; ALOX, lipoxygenases; CEA, carcino-embryonic antigen; KRAS, kirsten rat sarcoma viral oncogene; TNM, tumor-node-metastasis.