Progression and prognostic value of ECT2 in non-small-cell lung cancer and its correlation with PCNA

Figures & data

Table 1 Microarray databases detailing the mRNA expression of ECT2 (these databases were included in the meta-analysis)

First author	Year	Duration (months)	Histology	Stage	Patient number	Detection	Platform
Landi et al^Citation28	2008	NR	ADC	I–IV	58	Microarray	AffymetrixHgu133a
Kuner et al^Citation29	2009	NR	NSCLC	NR	58	Microarray	AffymetrixHgu133a
Baty et al^Citation30	2010	44.1	NSCLC	I–IV	56	Microarray	AffymetrixHgu133plus2.0
Takeuchi et al^Citation31	2006	109.1	NSCLC	I–III	158	Microarray	Agilent Homo sapiens 21.6K
Tomida et al^Citation32	2009	109.8	ADC	I–III	117	Microarray	Agilent-014850 G4112F
Zhu et al^Citation33	2010	111.5	NSCLC	I–II	133	Microarray	AffymetrixHgu133a
Hou et al^Citation34	2010	127.2	NSCLC	NR	91	Microarray	AffymetrixHgu133plus2.0
Lu et al^Citation35	2010	NR	NSCLC	I–IV	60	Microarray	AffymetrixHgu133plus2.0
Xie et al^Citation36	2011	82.44	NSCLC	I–III	55	Microarray	AffymetrixHgu133plus2.0
Rousseaux et al^Citation37	2013	140	NSCLC	I–IV	293	Microarray	AffymetrixHgu133plus2.0
Okayama et al^Citation38	2012	115.5	ADC	I–II	226	Microarray	AffymetrixHgu133plus2.0
Bild et al^Citation39	2006	81.2	NSCLC	NR	111	Microarray	AffymetrixHgu133plus2.0
Selamat et al^Citation40	2012	NR	ADC	I–IV	58	Microarray	IlluminaHumanWG-6v3.0
Botling et al^Citation41	2013	140	NSCLC	I–IV	196	Microarray	AffymetrixHgu133plus2.0
Tang et al^Citation42	2013	129.6	NSCLC	I–III	176	Microarray	IlluminaHumanWG-6v3.0
Tarca et al^Citation43	2013	NR	NSCLC	I–II	150	Microarray	AffymetrixHgu133plus2.0
Raponi et al^Citation44	2006	NR	SQC	NR	130	Microarray	AffymetrixHgu133a
Der et al^Citation45	2014	100.1	NSCLC	I–II	181	Microarray	AffymetrixHgu133plus2.0
Shedden et al^Citation46	2008	140	ADC	I–III	442	Microarray	AffymetrixHgu133a
Cancer Genome Atlas Research Network (CGARN)^Citation47	2015	NR	SQC	I–IV	135	Microarray	AffymetrixHgu133a
Lee et al^Citation48	2008	138.5	NSCLC	NR	138	Microarray	AffymetrixHgu133plus2.0

Abbreviations: ECT2, epithelial cell transforming sequence 2; NSCLC, non-small-cell lung cancer; ADC, adenocarcinoma; SQC, squamous cell lung cancer; NR, not reported.

Table 2 Correlation between ECT2 expression and the clinicopathological features of 202 lung cancer patients on TMAs (BC041115c and HlugA180Su05)

Variables	n	ECT2 expression		P-value
		≥6 (low expression)	>6 (high expression)
Age				0.648a
<60	105	65	40
≥60	97	57	40
Sex				0.571a
Female	71	41	30
Male	131	81	50
Differentiation				0.116a
Moderate + well	114	70	44
Poor	63	31	32
Tumor size				0.007b
T1 + T2	87	74	13
T3 + T4	23	13	10
Lymph node				0.011a
N^–	97	70	27
N^+	87	47	40
Stage				0.000a
I–II	124	92	32
III–IV	73	29	44

Notes:

a Pearson chi-squared test;

b Continuity correction chi-squared test; N^–, lymph node negative; N⁺, lymph node positive.

Abbreviations: ECT2, epithelial cell transforming sequence 2; TMAs, tissue microarrays.

Figure 1 ECT2 was enriched in various subtypes of lung cancer.

Notes: (A) Representative images of IHC staining of normal, squamous cell carcinoma, adenosquamous carcinoma, adenocarcinoma, bronchioloalveolar carcinoma, large cell carcinoma, small cell carcinoma, and atypical carcinoid; (B) quantitative IHC scores of ECT2 in various lung cancer subtypes; (C) meta-analysis of ECT2 mRNA expression in tumors vs normal; (D) meta-analysis of ECT2 mRNA expression in SQC vs ADC.

Abbreviations: ECT2, epithelial cell transforming sequence 2; IHC, immunohistochemical; T, tumor; N, normal; OR, odds ratio; ADC, adenocarcinoma; SQC, squamous cell lung cancer.

Figure 2 Protein level of ECT2 was associated with cancer progression and prognosis.

Notes: (A) Representative images and IHC scores of ECT2 in tumor vs matched adjacent tissue; (B) various stages; (C) various degrees of tumor cell differentiation; (D) various tumor sizes; (E) lymph node positive vs lymph node negative. N⁻, lymph node negative; N⁺, lymph node positive.

Abbreviations: ECT2, epithelial cell transforming sequence 2; IHC, immunohistochemical.

Figure 3 mRNA level of ECT2 was associated with cancer progression and prognosis.

Notes: (A) Meta-analysis of ECT2 mRNA expression at stage III + IV vs I + II; (B) stage III + IV vs I + II in ADC; (C) stage III + IV vs I + II in SQC; (D) tumor size T3 + T4 vs. T1 + T2; (E) lymph node positive vs lymph node negative; (F) the mRNA expression of ECT2 at different degrees of tumor cell differentiation in GSE68465. N⁻, lymph node negative; N⁺, lymph node positive.

Abbreviations: ECT2, epithelial cell transforming sequence 2; IHC, immunohistochemical staining; CGARN, Cancer Genome Atlas Research Network; OR, odds ratio; ADC, adenocarcinoma; SQC, squamous cell lung cancer.

Table 3 ECT2 expression in ADC is an independent prognostic factor for ADC patients (HlugA180Su05)

Variables	Univariate analysis		Variable selection
	HR (95% CI)	P-value	HR (95% CI)	P-value
Sex (female/male)	0.733 (0.457–1.176)	0.198
Age (>60/≥60)	1.011 (0.632–1.619)	0.962
Differentiation (poor/moderate + well)	2.360 (0.859–6.486)	0.096
Stage (II + III/I)	2.658 (1.483–4.765)	0.001
Tumor size (T3 + T4/T1 + T2)	1.424 (0.838–2.419)	0.191
Lymph node metastasis (N^+/N^−)	2.672 (1.541–4.633)	0.000	2.847 (1.604–5.054)	0.000
ECT2 expression (high/low)	2.010 (1.204–3.355)	0.008	2.464 (1.375–4.418)	0.002

Notes: N⁻, lymph node negative; N⁺, lymph node positive.

Abbreviations: ECT2, epithelial cell transforming sequence 2; ADC, adenocarcinoma; HR, hazard ratio.

Figure 4 Overexpression of ECT2 protein predicted poor survival.

Notes: Kaplan–Meier curve of patients with low or high ECT2 expression in ADC. Median level of IHC scores was used to divide patients into subgroups;

Abbreviations: ECT2, epithelial cell transforming sequence 2; HR, hazard ratio; ADC, adenocarcinoma; IHC, immunohistochemical staining.

Figure 5 ECT2 expression correlated with PCNA expression.

Notes: (A) Representative images of IHC staining of PCNA; (B) representative images of IHC staining of PCNA and ECT2 in the same lung cancer tissues; (C) IHC scores of PCNA in lung cancer; (D) the correlation between ECT2 and PCNA based on protein expression; (E) based on the mRNA level of the GSE4342 cell line; (F) GSE32474 cell line; (G) GSE31210 cell line; (H) GSE68465 cell line; (I) GSE30219 cell line. N⁻, lymph node negative; N⁺, lymph node positive.

Abbreviations: IHC, immunohistochemical staining; PCNA, proliferating cell nuclear antigen; ECT2, epithelial cell transforming sequence 2; HR, hazard ratio; ADC, adenocarcinoma; SQC, squamous cell lung cancer.

Figure S1 Overexpression of ECT2 mRNA predicted poor survival.

Notes: Kaplan–Meier curve of patients with low or high ECT2 expression. Median of mRNA expression was used to divide patients into subgroups. (A) Overall survival; (B) progression-free survival; (C) overall survival in ADC; (D) progression-free survival in ADC; (E) overall survival in SQC; (F) progression-free survival in SQC.

Abbreviations: ECT2, epithelial cell transforming sequence; HR, hazard ratio; ADC, adenocarcinoma; SQC, squamous cell lung cancer.

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