Circulating Tie2-Expressing Monocytes: A Potential Biomarker for Cervical Cancer

Figures & data

Figure 1 TEMs increased in peripheral blood of CC patients. (A) Detection of cTEMs in the peripheral blood (flow cytometric analysis). (B) Representative images of immunofluorescence assays for CD14 (green) and Tie2 (red) expression in TEMs and TNegMs which were isolated by FACS. Cell nuclei were labeled with DAPI (blue). Both CD14 (green) and Tie2 (red) were expressed in TEMs, while only CD14 (green) was expressed in TNegMs. Scale bar, 20 μm. (C) The percentage of cTEMs (Tie⁺CD14⁺) in total CD14⁺ monocytes was detected in peripheral blood of 7 HV, 17 UF, 24 CIN II, 31 CIN III and 99 CC patients. (D) The percentage of cTEMs is significantly increased in CC patients (Mann–Whitney u-test). *P<0.05, ** P<0.01 and *** P<0.001. (E) Boxplot of the Jonckheere–Terpstra test for the trend of percentage of cTEMs changing with the severity of cervical lesions. P<0.001. (F) Representative images of immunofluorescence assays for CD14 (green) and Tie2 (red) expression in PBMCs of HV and CC patient. Cell nuclei were labeled with DAPI (blue). Scale bar, 20 μm.

Abbreviations: CC, cervical cancer; HV, healthy volunteers; UF, uterine fibroid patients.

Figure 2 The diagnostic value of cTEM for cervical lesions. ROC analyses were performed in order to assess the diagnostic value of percentage of cTEMs for differentiating non-CC (HV, UF and CIN patients) from CC patients (A), HV, UF and CIN II from CIN III and CC patients (B), and for differentiating HV and UF from CIN and CC patients (C).

Table 1 The Relation of Circulating TEMs to Clinical-Pathological Parameters in Cervical Cancer

Clinicopathological Parameters	N	TEMs % Median (Range)	Mann–Whitney u-Test P value
Age (years)
<45	27	7.01 (3.23~11.43)	0.37
≥45	72	6.04 (3.56~13.78)
FIGO stage
IA1-IB1	51	5.13 (3.23~8.83)	<0.0001
IB2-IIB	48	7.75 (4.71~13.78)
Histological type
Squamous cell carcinoma	80	6.42 (3.23~13.78)	0.12
Adenocarcinoma	19	5.22 (4.02~9.13)
Tumor size (cm)
≤2	38	5.04 (3.23~8.56)	<0.0001
>2	61	7.24 (4.02~13.78)
Deep (outer third) stromal invasion
Negative	52	5.16 (3.23~8.83)	<0.0001
Positive	47	7.81 (4.71~13.78)
Histologic grade
Well/Moderately differentiated	72	6.11 (3.56~13.78)	0.89
Poorly differentiated	27	6.83 (3.23~10.64)
Parametrial invasion
Negative	84	6.02 (3.23~10.64)	<0.0001
Positive	15	8.83 (4.73~13.78)
Lymph-vascular space invasion
Negative	53	5.33 (3.23~10.64)	<0.0001
Positive	46	7.65 (4.57~13.78)
Lymph node metastasis
Negative	75	5.58 (3.23~9.68)	<0.0001
Positive	24	8.73 (6.28~13.78)

Abbreviations: TEMs, Tie2-expressing monocytes; FIGO, International Federation of Gynecology and Obstetrics.

Figure 3 Percentage of cTEMs is correlated with CC progression. High level of cTEMs were significantly associated with FIGO stage (A), tumor size (B), deep stromal invasion (C), parametrial invasion (D), lymph-vascular space invasion (E) and LNM (F). ***P<0.001.

Table 2 Univariate Analysis and Multivariate Analysis of Lymph Node Metastasis in Patients with Cervical Cancer (N=99)

Parameters	Univariate Analysis			Multivariate Analysis
	OR	95% CI	P	OR	95% CI	P
Age (years), ≥45 vs. <45	0.48	0.18~1.30	0.150	–	–	–
Histological type, adenocarcinoma vs. squamous cell carcinoma	3.04	0.64~14.22	0.161	–	–	–
FIGO stage, IB2-IIB vs. IA1-IB1	7.28	2.26~23.52	0.001	1.18	0.21~6.61	0.854
Deep stromal invasion, positive vs. negative	5.83	1.96~17.42	0.002	1.64	0.41~6.59	0.487
Lymph-vascular space invasion, positive vs. negative	8.62	2.66~27.95	<0.001	1.46	0.33~6.54	0.621
Tumor size (cm), >2 vs. ≤2	8.50	1.86~38.84	0.006	3.51	0.44~28.29	0.238
Parametrial invasion, positive vs. negative	10.92	3.21~37.21	<0.001	5.94	1.19~29.58	0.030
TEMs %, ≥7 vs. <7	20.00	5.34~74.86	<0.001	6.25	1.18~33.00	0.031

Abbreviations: OR, odds ratio; CI, confidence interval.

Figure 4 Percentage of cTEMs is an independent predictor of CC LNM.