The Measurement Of Peripheral CECs And V-CECs In Patients With Endometrial Carcinoma And Clinical Significance

Objective: Tumor angiogenesis has a close relationship with the occurrence and development of tumor. Studies have shown that the circulating endothelial cells(CECs) form lumen by proliferation, migration, adhesion and take part in tumor angiogenesis, which provides not only oxygen and nutrients for the tumor growth, but also the path for tumor metastasis and development. CECs are further divided into resting circulating endothelial cells(r CECs), viable circulating endothelial cells(V-CECs) and apoptotic CECs, while V-CECs play the most important role. It was reported that the level of CECs increased obviously in the peripheral blood of advanced tumor patient, while the level of CECs of stable tumor patient was similar with the level of the healthy people. It was also found that the number of CECs was decreased obviously in non-small cell lung cancer patients who got clinical benefit after treatment with chemotherapy and endostar. These studies showed that CECs and V-CECs could be used as biomarkers to evaluate the level of tumor angiogenesis and the efficacy of the drugs of anti-tumor angiogenesis. The aim of the study is to investigate the levels of CECs and V-CECs in the peripheral blood of the endometrial carcinoma patients and healthy women, and to explore its clinical significance.Methods: Based on the case-control study, blood samples were collected from 58 cases and 20 controls. CECs were measured by the methods of Hladovec and flow cytometry(FCM), meanwhile V-CECs were detected by flow cytometry. The definition of CECs was CD45-CD146+ cells, while the definition of V-CECs was CD45-CD146+CD106+ cells, according to literature.All results were analyzed by applying SPSS software(version 13.0). If the distribution of data was normal, the results were expressed as x ±s. Comparison of two groups was performed by the t-test. Comparison of multiple groups was performed by analysis of variance. If the distribution of data was abnormal, the results of experiment were expressed as median, performed by the nonparametric test. The correlation between different indicators was performed by Spearman Correlation. Statistical differences with a P value < 0.05 were considered significant.Results: 1 Comparison between the cases and the controls 1.1 Comparison of the data of CECs, V-CECs between the cases and the controlsThe distributions of data were normal, expressed as x ±s. The results by Hladovec method showed that CECs level in the case group(18.647±9.550/μL) were markedly higher than that of the controls(5.125±3.191/μL), the difference was statistical significance(P<0.001). The results by flow cytometry showed that the levels of CECs, V-CECs in the case group[( 2.340±1.541)%,(1.090±0.721)%] were obviously higher than that of the controls[(0.765±0.532)%,( 0.236±0.194) % ], the differences were statistical significance(P< 0. 001,P< 0. 001).1.2 Comparison of the ratio of V-CECs/CECs between the cases and the controlsThe distributions of the data were normal, expressed as x ±s. the level of V-CECs/CECs in the case group(47.630±13.770)% was obviously higher than that of the controls(27.801±11.429)%, the difference was statistical significance(P< 0. 001).2 Comparison of the data of CECs, V-CECs between the cases 2.1The relationship between the levels of CECs, V-CECs and different stagesAccording to different stages, the case group could be divided into the groups of phase I、phase II and phase III、IV(namely the advanced group). The distributions of data were abnormal, then expressed as median.The results by Hladovec method showed that the CECs level of the phase I(20.00/μL)was higher than that of phase II and advanced phase(18.75/μL,19.50/μL), but the difference was not statistically significant(P=0.808).The results by flow cytometry showed that the CECs levels of the groups of phase I、phase II(2.47%,2.19%)were higher than that of the advanced group(1.99%), while the V-CECs levels of the groups of phase I、phase II(1.11%,0.90%) were lower than that of the advanced group(1.14%), but all the differences were not statistically significant(P=0.813,P=0.922).2.2 Relationship between the levels of CECs, V-CECs and different myometrial invasionAccording to different myometrial invasion, the case group could be divided into the groups of shallow and deep. The distributions of the data were normal, expressed as x ±s.The results by Hladovec method showed that the CECs level of the shallow group( 18.904±9.098/μL) was higher than that of deep group(16.417±13.738/μL), but the difference was not statistically significant(P=0.682).The results by flow cytometry showed that the levels of CECs, V-CECs in the shallow group [(2.363±1.495)%,(1.092±0.697)% ] were higher than that of deep group [(2.138±2.057)%,(1.068±0.988)% ], but the differences were not statistically significant(P=0.738,0.940).2.3 Relationship between the levels of CECs, V-CECs and different pathological typesAccording to different pathological types, the case group could be divided into the group of adenocarcinoma and squamous carcinoma and the group of special pathological types(including clear cell carcinoma and serous papillary adenocarcinoma). The distributions of data were abnormal, expressed as median.The results by Hladovec method showed that the CECs level of the group of adenocarcinoma and squamous carcinoma(20.00/μL)was higher than the group of special pathological types(17.50/μL), but the difference was not statistically significant(P=0.806).The results by flow cytometry showed that the CECs, V-CECs levels of the group of adenocarcinoma and squamous carcinomawere were lower than that of the group of special pathological types, but all the differences were not statistically significant(P=0.591,P=0.480).2.4 Relationship between the levels of CECs, V-CECs and the cell differentiationAccording to the cell differentiation, the case group could be divided into the groups of grade I、II、III.The results by Hladovec method was shown abnormally distributed, expressed as median. The CECs levels of the groups of grade I、II(19.50/μL,20.00/μL)were higher than that of grade III(18.50/μL), but the differences were not statistical significance(P=0.872).The results by flow cytometry were differently distributed. The distribution of the data of CECs was normal, expressed as x ±s. The level of CECs in the group of grade I、II[(2.166±1.229)%,(2.355±1.765)%]was lower than that of grade III(2.580±1.533)%, but the differences were not statistically significant(P=0.772). The distribution of the data of V-CECs was abnormal, expressed as median. The levels of V-CECs in the group of grade I、II(1.00%,1.01%)were lower than that of grade III(1.145%), but the differences were not statistically significant(P=0.557).2.5 Relationship between the levels of CECs, V-CECs and the maximum diameter of tumorAccording to the maximum diameter of tumor, the case group could be divided into the groups of <2cm and ≥2cm. The distributions of CECs, V-CECs were normal, expressed as x ±s.The results by Hladovec method showed that the CECs level in the <2cm group( 16.797±9.501/μL) was markedly lower than that of the ≥2cm group(21.905±8.941/μL). The difference was statistically significant(P=0.049).The results by flow cytometry showed that the levels of CECs, V-CECs in the <2cm group[(1.984±1.383)%,(0.899±0.644)% ] were obviously lower than that of the≥2cm group[(2.967±1.636)%,(1.426±0.742)% ]. The differences were statistically significant(P=0.018, P=0.006).3 Relationship between the levels of CECs, V-CECs and the value of CA125The Spearman Correlation between CECs measured by Hladovec and the value of CA125 was 0.075. There was no statistical significance(P=0.576).The Spearman Correlations between CECs, V-CECs measured by flow cytometry and the value of CA125 were 0.139、0.155. There were no statistical significance(P=0.297,0.246).Conclusion: 1 The levels of CECs, V-CECs and the ratio of V-CECs/CECs were markedly elevated in the endometrial carcinoma patients than that in the healthy women, indicating that CECs, V-CECs may be used as biomarkers to evaluate the level of tumor angiogenesis and the efficacy of drugs of anti-tumor angiogenesis.2 The levels of CECs, V-CECs have no relationship with the stage、myometrial invasion、pathological type、cell differentiation of endometrial carcinoma.3 The levels of CECs, V-CECs have a close relationship with the maximum diameter of endometrial carcinoma.4 There were no obvious correlations between the levels of CECs, V-CECs and the value of CA125.