Loss of Cirbp expression is correlated with the malignant progression and poor prognosis in nasopharyngeal carcinoma

Figures & data

Figure 1 Cirbp was downregulated in the clinical tissue specimens of nasopharyngeal carcinoma (NPC). (A) Cirbp expression in the indicated 8 NPC cell lines were measured by qRT-PCR. (B) The levels of Cirbp transcript were examined in 38 NPC clinical specimens and 23 non-cancerous nasopharyngeal (NP) biopsies by qRT-PCR. (C,D,E) The expression levels of Cirbp in NPC clinical specimens and non-cancerous nasopharyngeal biopsies from GEO datasets. (F) Cirbp expression in NPC and non-cancerous nasopharyngeal epithelial tissues based on IHC. a and b: High expression of Cirbp in non-cancerous nasopharyngeal epithelial tissues; c and d: low Cirbp expression in NPC tissue biopsies; e and f: high Cirbp expression in NPC tissue biopsies. The brown staining shows Cirbp immunoreactivity. (G) Cirbp expression was markedly lower in the NPC tissue biopsies than that in the non-cancerous nasopharyngeal epithelial tissues.

Table 1 Expression of Cirbp in 61 non-cancerous nasopharyngeal (NP) epithelial tissues and 177 nasopharyngeal carcinoma (NPC) tissues

Variables	n	Cirbp expression		χ2	P
		Low, n (%)	High, n (%)
Non-cancerous NP epithelial tissues	61	0 (0.0)	61 (100.0)	54.491	<0.001
NPC	177	95 (53.7)	82 (46.3)

Figure 2 Cirbp downregulation was associated with the tumor malignant progression in NPC patients. (A) Representative pictures of Cirbp expression in nasopharyngeal carcinoma (NPC) clinical biopsies of different TNM stages, clinical stage and histological type. High expression of Cirbp was detected in the T1 (a), N0 (c), M0 (e) and I (g) stages, and differentiated nonkeratinizing carcinoma (DNKC)(i) of NPC biopsies, while low expression of Cirbp was observed in the T3 (b), N2 (d), M1 (f) and IV (h) stages, and undifferentiated carcinoma (UDC)(j) of tumor. (B) The number and percentage of low and high expression of Cirbp based on different clinicopathological features. **P<0.01 and ^#P<0.001. 

Table 2 Association between the clinicopathological features and Cirbp expression in 177 nasopharyngeal carcinoma (NPC) tissue specimens

Characteristics	Case No. (n)	Cirbp expression		χ2	P
		High, n (%)	Low, n (%)
Sex
Female	46	20 (43.5)	26 (56.5)	0.078	0.781
Male	131	62 (47.3)	69 (52.7)
Age (years)
＜50	82	37 (45.1)	45 (54.9)	0.022	0.883
≥50	95	45 (47.4)	50 (52.6)
Histological type
DNKC	28	21 (75.0)	7 (25.0)	9.670	0.002
UDC	149	61 (40.9)	88 (59.1)
T classification
T1-T2	118	68 (57.6)	50 (42.3)	16.839	＜0.001
T3-T4	59	14 (27.1)	45 (72.9)
N classification
N0-N1	116	65 (56.0)	51 (44.0)	11.647	＜0.001
N2-N3	61	17 (27.9)	44 (72.1)
M classification
M0	141	75 (53.2)	66 (46.8)	11.813	0.001
M1	36	7 (19.4)	29 (80.6)
Clinical stage
I - II	59	40 (67.8)	19 (32.2)	15.135	＜0.001
III - IV	118	42 (35.6)	76 (64.4)

Abbreviations: DNKC, differentiated nonkeratinizing carcinoma; UDC, undifferentiated carcinoma.

Table 3 Univariate and multivariate analyses of the overall survival of 177 patients with nasopharyngeal carcinoma (NPC)

Variables	Univariate analysis		Multivariate analysis
	HR (95% CI)	P	HR (95% CI)	P
Gender
Female vs Male	1.747 (0.837–3.646)	0.137	2.355 (0.878–6.312)	0.089
Age (y)
＜50 vs ≥50	0.770 (0.427–1.390)	0.386	2.565 (1.197–5.497)	0.015
Histological type
DNKC vs UDC	2.150 (0.941–4.909)	0.046	6.588 (2.462–17.631)	0.000
T classification
T1-T2 vs T3-T4	0.474 (0.253–0.887)	0.020	0.582 (0.229–1.484)	0.257
N classification
N0-N1 vs N2-N3	0.475 (0.256–0.881)	0.018	0.291 (0.118–0.721)	0.008
M classification
No vs Yes	0.146 (0.067–0.318)	<0.001	0.11 (0.04–0.299)	0.000
Clinical stage
Ⅰ-Ⅱ vs Ⅲ-Ⅳ	0.481 (0.255–0.907)	0.024	0.228 (0.077–0.677)	0.008
Cirbp expression
High vs Low	2.196 (1.218–3.960)	0.009	5.388 (2.474–11.734)	0.000

Abbreviations: HR, hazard ratio; 95% CI, 95% confidence interval; DNKC, differentiated nonkeratinizing carcinoma; UDC, undifferentiated carcinoma. 

Table 4 Correlation between Cirbp and E-cadherin & Ki6 in 177 nasopharyngeal carcinoma (NPC) tissue specimens

Markers	Case no.	Cirbp expression		r	P
		High, n (%)	Low, n (%)
E-cadherin
High	88	57 (64.8)	31 (35.2)	0.368	＜0.001
Low	89	25 (28.1)	64 (71.9)
Ki67
＜30	78	52 (66.7)	26 (33.3)	0.371	＜0.001
≥30	102	30 (29.4)	72 (70.6)

Note: Spearman correlation.

Figure 3 Cumulative Kaplan–Meier overall survival curves of 177 nasopharyngeal carcinoma (NPC) patients. (A) Survival curves for NPC patients with DNKC and UDC. (B-E) Survival curves for NPC patients according to TNM staging and clinical stage. (F) Cumulative overall survival curves of 177 NPC patients with different Cirbp expression (high or low). The P-value was calculated by log-rank test.

Abbreviations: DNKC, differentiated nonkeratinizing carcinoma; UDC, undifferentiated carcinoma. 

Figure 4 Association between Cirbp expression and E-cadherin expression or proliferation index Ki67. (A) Representative pictures of IHC staining of Cirbp, E-cadherin and Ki67 in nasopharyngeal carcinoma (NPC) tissue biopsies. (B) The number and percentage of cases with high or low expression of the indicated genes. (C) The expression of Cirbp and Ki67 in NP69 and NPC cancer cell lines was examined by qRT-PCR. (D) The expression of Cirbp and E-cadherin in NP69 and NPC cancer cell lines was examined by qRT-PCR. *P<0.05, **P<0.01 and ^#P<0.001.