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Amiraslanov Ahliman Tapdig, Academician, Chief of Department of Oncology, Azerbaijan Medical University Gaziyev Abuzer Yusif, professor, doctor of medical sciences, Department of Oncology, Azerbaijan Medical University
Safarova Indira Adil, doctoral student of Oncology Department of Azerbaijan Medical University Mehdiyeva Nigar ismail, associate professor, Department of Oncology, Azerbaijan Medical University Department of Oncology of Azerbaijan Medical University E-mail: [email protected]
DIAGNOSTIC SIGNIFICANCE OF ENDOTHELIN-1 AND L-FABP IN PATIENTS WITH CERVICAL CANCER
Abstract: In this article, the diagnostic significance of endothelin-1 and L-FABP in diagnostic of cervical cancer (CC)has been studied. To realize the goal, venous blood of 53 patients with CC and 25 patients with cervical leuko-plagia (CL) was examined. The results of the study showed that in patients with cervical cancer, the level of endothelin-1 and L-FABP were higher by 38.5%, vs to control and by 14.6%, compared with the results of patients with CL. To clarify the significance of endothelin-1 and L-FABP in the development of CC, we studied the content of some tumor markers (CA125, PEA, CA 19-9, AFP and ^-CG) and their correlation. There were statistically significant increases in CA125, PEA, CA 19-9, AFP and ^-CG in CC patients, compared with the data of patients with CL.
Keywords: cervical cancer, leukoplagia, endothelin-1, L-FABP and oncomarkers.
Cervical cancer (CC) is one of the most common form of patients with advanced disease. False positive results have of neoplasm among the women malignancies. Each year, been noted in some disorders, both malignant and benign, 371.000 of new cases of cervical cancer are registered in the which to some extent reduce the specificity and sensitivity of world and 190.000 of women die each year [6; 7; 13]. Early the marker. Therefore, the screening of CC includesCA125, detection and treatment of precancerous diseases of the cervix CEA, CA 19-9, AFP and beta CG [3; 4]. can completely prevent the cancer. The use of new instrumen- Recently, much attention has been paid to the role of bio-
tal diagnostic methods is not always available and sometimes chemical markers in the pathogenesis of CC. Endothelin-1 and there are some difficulties for clarifying the diagnosis [1]. Dis- L-FABP belong to the number of biological active peptides, easerelated mortality increases because of the lack of a reliable which play an important role in carcinogenesis. Endothelin-1 screening technique for early CC detection. Therefore, the is a polypeptide of 21 amino acid residues, has vasoconstrictor development of highly informative methods for the detection and mitogenic activity. Endothelin-1 is the one of the most sig-of tumors of the ovaries and cervix, as well as early diagnosis nificant regulators ofthe vascular endothelium function besides, of recurrence of the disease is one of the most important tasks by controlling mitogenesis, cell survival, angiogenesis, bone re-of oncogynecology. modeling, stimulation of tumor-permeating immune cells, ep-
In recent years investigation of tumor markers' (CA 125, ithelial-mesenchymal changes play an important role in tumor CEA, CA 19-9, AFP and ^-CG) level in blood serum takes a development and metastasis [8; 9; 17]. L-FABP is a member great place in CC diagnostic [4]. of the fatty acid binding group that is involved in intracellular
Tumor markers are important indicators of CC clinical transfer of biologically active fatty acids and is involved in in-presentation. Due to its high sensitivity and specificity, CA125 tracellular signaling pathways, cell growth and differentiation. has a clearly defined and confirmed role in the monitoring of In addition, by preventing the increase in the concentrations of cervical carcinoma. It rises in approximately 50% of malig- intracellular fatty acids, protect cells from their cytotoxic effects. nant diseases of female reproductive organs and in 75-90% Using a series of studies on the effects of chemical carcinogens
on rat liver, the participation of L-FABP in cell growth and differentiation was determined. A significant increase in L-FABP was observed in normal as well as carcinogen-induced hepato-cytes. In addition to acting as a fatty acid receptor, L-FABP is the target protein for several genotoxic carcinogens. The binding of these carcinogens to L-FABP promotes mitogenesis and tumor development [5; 11; 14].
The aim of this work was a comparative study of the diagnostic significance of endothelin-1 and L-FABP in blood plasma in patients with CC and cervical leukoplagia (CL).
Material and methods. The content of endothelin-1, L-FABP and some oncomarkers (CA125, PEA, CA 19-9, AFP and P-CG) was studied in patients with cervical cancer who were on treatment at the oncological clinic of the Azerbaijan Medical University. The main group consisted of 53 patients with cervical cancer (mean age - 50.3 ± 1.3 years). The comparison group included 25 women with CL (mean age 42.4 ± ± 2.4 years). The control group consisted of 12 practically healthy women (mean age - 36.9 ± 2.9 years).
In 7(28.0%) patients a simple form of CL was detected, in 13(52.0%) - proliferative, and in 5(20.0%) - atypical. Among
the patients with cervical cancer, 2(3.8%) had a squamous cell cornifiredtype, 43(81.11%) had a squamous cell noncor-nifired type, 6(11.9%) had an adenocarcinoma, and 2(3.8%) had another types.
The concentration of endothelin 1, L-FABP, CA125, PEA, CA 19-9, AFP and ^-CG was determined in blood serum in the study groups using Vector Best immunoassay test systems (Russia), on the STAT FAX immunoassay analyzer. These indicators were determined at the time of admission to the hospital before the start of surgical treatment.
The results were represented as M ± m, where M is the arithmetic mean, m is the error of the mean. The reliability of the differences between the groups was assessed using the Mann-Whitney test, for related values or the Student's t-test for unbound values. The difference was considered significant for p < 0.05. The correlation analysis was carried out according to the method of Spearman.
Results and discussion:
As can be seen from the obtained results, the content of endothelin-1 and L-FABP in the blood serum increases in both groups of patients (Table 1).
Table 1. - Dispersion analysis of the studied parameters in patients with cervical cancer
Data Patients group N Overage Stand. deviat. Stand. error 95% Hgr 95% Lgr Fiser P
1 2 3 4 5 6 7 8 9 10
L-FABP, ng/ ml Squamous cellcornifired 2 2.41 2.96 2.09 0 28.97 5.385 .003
Squamous cellnoncor-nifired 43 2.00 1.42 0.22 1.56 2.44
Adenocarcinoma 6 3.80 0.17 0.07 3.62 3.97
The others 2 5.10 2.55 1.80 0 27.97
Total 53 2.34 1.60 0.22 1.89 2.78
Endothelin, pg/m1, Squamous cellcornifired 2 9.0 0.1 0.0 8.3 9.6 11.333 .000
Squamous cellnoncor-nifired 43 8.5 0.6 0.1 8.3 8.7
Adenocarcinoma 6 9.7 0.3 0.1 9.4 10.1
The others 2 10.3 0.4 0.3 7.1 13.4
Total 53 8.7 0.8 0.1 8.5 9.0
CEA, pg/ml Squamous cellcornifired 2 7.30 0.99 0.70 0 16.19 15.631 0.001
Squamous cellnoncor-nifired 43 5.36 1.62 0.25 4.86 5.86
Adenocarcinoma 6 7.78 1.34 0.55 6.38 9.19
The others 2 8.10 0.57 0.40 3.02 13.18
Total 53 5.81 1.79 0.25 5.32 6.31
CA125, ng/m Squamous cellcornifired 2 36.75 17.32 12.25 0 192.40 4.187 0.010
Squamous cellnoncor-nifired 43 34.81 12.02 1.83 31.11 38.51
Adenocarcinoma 6 50.30 2.37 0.97 47.81 52.79
1 2 3 4 5 6 7 8 9 10
CA125, ng/ml The others 2 50.70 1.70 1.20 35.45 65.95
Total 53 37.24 12.43 1.71 33.81 40.66
Squamous cellcornifired 2 27.3 11.3 8.0 0 128.9 1.270 0.295
CA19-9, ng/ Squamous cellnoncor-nifired 43 39.0 11.7 1.8 35.4 42.6
ml Adenocarcinoma 6 35.7 15.0 6.1 19.9 51.5
The others 2 49.4 1.7 1.2 34.2 64.6
Total 53 38.6 12.1 1.7 35.3 41.9
Squamous cellcornifired 2 10.3 4.0 2.9 0 46.5 0.950 0.424
AFP, pg/ml Squamous cellnoncor-nifired 43 10.4 2.4 0.4 9.6 11.1
Adenocarcinoma 6 11.8 3.2 1.3 8.5 15.1
The others 2 12.4 0.2 0.2 10.4 14.3
Total 53 10.6 2.5 0.3 9.9 11.3
Squamous cellcornifired 2 4.65 0.92 0.65 0 12.91 6.199 0.001
ß-CG pg/ml Squamous cellnoncor-nifired 43 6.29 1.47 0.22 5.83 6.74
Adenocarcinoma 6 8.93 2.61 1.07 6.19 11.67
The others 2 7.90 0.71 0.50 1.55 14.25
Total 53 6.58 1.83 0.25 6.08 7.09
In our investigation the level ofendothelin- 1and L -FABP in patients with CC increases by 38.5%(8.7 ± 0.1 pg/ml, p < 0.001) or in 5.2 times (2.34 ± 0.22 pg/ml, p < 0.001). in CL patients it was 20.9%(7.62 ± 0.17 pg/ml, p < 0.05) or in 1.7 times higher than in control cohort (0.76 ± 0.06; p < 0.01).A comparative analysis of the results showed an increase in en-dothelin-1 and L-FABP level in patients with CC by 14.6% (p < 0.001) or 3.1 times (p < 0.001) compared to those of CL patients. The highest concentration of these markers was observed in the group of patients with cervical adenocarcinoma (9.7 ± 0.1 pg/ml and 3.80 ± 0.07 pg/ml, respectively) and in patients with atypical forms of CL (8.00 ± 0.64 pg/ml and 1.03 ± 0.03 pg/ml, respectively).
Today, endothelin-1 is considered to be a marker of metastasis, angiogenesis and a predictor of the severity of the course of various neoplasms. Since endothelin acts predominantly locally, it is natural to assume that an active synthesis and entry into the bloodstream of the tumor site may be the cause of the progression of angiogenesis and aggravation of
the severity of the course. It is known that in tissue damage the endothelin system reacts first, so a sharp increase in endothelin-1 plasma level is considered to be a marker of the activity of the destruction process, including tumor formation. In addition, tumors also contain cells of the immune system that release factors such as endothelin, prostaglandins and tumor necrosis factor alpha (TNF-a) [2; 8; 9; 17].
Increasing the expression of L-FABP leads to the development and progression of the tumor and can also serve as a useful diagnostic marker [5; 10; 14].
In order to study the severity of the oncological process, various cancer markers were determined in all patients. According to the obtained results, the content of CEA increased in CC (5.81 ± 0.25 ng/ml, p < 0.001) and in patients with CL (3.70 ± 0.29 ng/ml; p < 0.01) compare with the control group (CEA - 2.09 ± 0.36 ng/ml). Cervical adenocarcinomas (7.78 ± ± 0.55 ng/ml) and an atypical form of CL (4.98 ± 0.47 ng/ml) showed, the greatest levels of CEA (table 2).
Data Patients group N Overage Stan. deviat Stand. error. 95% Hgr 95% Lgr Fiser P
1 2 3 4 5 6 7 8 9 10
L-FABP, ng/ml Simple 7 0.43 0.28 0.11 0.17 0.70 17.159 0.000
Proliferative 13 0.82 0.14 0.04 0.74 0.91
Atypical 5 1.03 0.08 0.03 0.94 1.13
Total 25 0.76 0.28 0.06 0.64 0.87
Table 2.- Dispersion analysis of the studied parameters in patients with CL
1 2 3 4 5 6 7 8 9 10
Endothelin, pg/ml, Simple 7 7.00 0.44 0.17 6.59 7.41 3.219 0.059
Proliferative 13 7.82 0.58 0.16 7.47 8.16
Atypical 5 8.00 1.43 0.64 6.22 9.78
Total 25 7.62 0.85 0.17 7.27 7.97
CEA, pg/ml Simple 7 2.66 1.44 0.54 1.33 3.99 5.279 0.013
Proliferative 13 3.76 1.17 0.32 3.06 4.47
Atypical 5 4.98 1.04 0.47 3.69 6.27
Total 25 3.70 1.43 0.29 3.11 4.29
CA125, ng/ml Simple 7 8.3 1.0 0.4 7.4 9.2 29.381 0.000
Proliferative 13 15.7 3.9 1.1 13.3 18.0
Atypical 5 25.3 5.6 2.5 18.3 32.3
Total 25 15.5 6.9 1.4 12.7 18.4
CA19-9, ng/ ml Simple 7 13.1 6.0 2.3 7.6 18.7 2.616 0.096
Proliferative 13 19.0 4.7 1.3 16.2 21.9
Atypical 5 18.7 7.5 3.4 9.3 28.0
Total 25 17.3 6.1 1.2 14.8 19.8
AFP, pg/ml Simple 7 5.01 2.16 0.81 3.02 7.01 0.005 0.995
Proliferative 13 4.95 1.52 0.42 4.04 5.87
Atypical 5 5.04 1.80 0.80 2.81 7.27
Total 25 4.99 1.69 0.34 4.29 5.68
ß-CG, pg/ml Simple 7 2.89 1.85 0.70 1.17 4.60 0.860 0.437
Proliferative 13 2.36 0.27 0.07 2.20 2.52
Atypical 5 2.90 0.51 0.23 2.27 3.53
Total 25 2.62 1.00 0.20 2.20 3.03
It is known that the level of CEA correlates with the grade of the cancer. It is shown that low grade tumors produce CEA more actively. According to numerous authors, the marker has prognostic significance, which based on the fact that a high initial level of CEA in the serum indicates a high risk of early relapse [4; 16].
CA 125 is a glycoprotein produced by cells of malignant serous ovarian tumors. It is one of the important tumor markers for monitoring the severity of clinicand effectiveness of the therapy of various types of ovarian cancer (serous, endometrial, clear cell) and CC. CA 125 testing can detect a relapse of the disease 3-4 months before its clinical manifestation [3; 4].
According to our results patients with CC had significantly high level of CA 125. Thus the CA125 concentration in patients with CC was 37.24 ± 1.71 ng/ml(p < 0.001) and in patients with CL 15.5 ± 1.4 ng/ml(p < 0.01). As seen from the results, the level of CEA and CA 125 in patients with cervical cancerwas higher by 57.3%(p < 0.001) and 2.4 times (p < 0.001), respectively compared to the results of patients with CL.
The results of the analysis showed that the content of CA 19-9 in patients with cervical cancer was 3.1 times higher than in the control 38.6 ± 1.7 ng/ml (p < 0.001) CL patients have
about 40% increase of CA19-9 level 17.3 ± 1.2 (p < 0.05) with respect to the control values (12.5 ± 1.8 ng/ml). The concentration of CA19-9 was also different in patients with adenocarcinoma of cervix (35.7 ± 6.1 ng/ml) and in patients with atypical form of CL (19.0 ± 1.3 ng/ml).
in our investigation the AFP content was 3.2 times higher than in control 10.6 ± 0.3 ng/ml(p < 0.001) and 3.28 ± 0.49 ng/ml respectively as can be seen, the increased amountof AFPwas detected in patients with CL (52.3%) 4.99 ± 0.34 ng/ml (p < 0.01). The average concentration of AFP in patients with cervical adenocarcinoma was 11.8 ± 1.3 ng/ml, and in patients with atypical form of CL it was, 5.04 ± 0.80 ng/ml. Consequently we can say that increase of AFP in the blood ofCCpatients indicates the predisposition to the high risk of relapse. It is generally believed that the malignant tumors of liver and ovaries, as well as the cervix produce AFP that enters the bloodstream. Tumor cells of the ovaries and cervix also contain elements of syncytiotrophoblast, therefore they synthesize HG along with AFP [15].
In patients with cervical cancer, the level of |?-HCG increased by 3.9 times (6.58 ± 0.25 ng/ml, p < 0.001) as compared to the normal. In patients with CL, the concentration
of |-HCG was 2.62 ± 0.20 ng/ml, which is 54.6% higher than in the control group (1.69 ± 0.36 ng/ml, p < 0.05). As can be seen from the results, the level of |-HCG in the adenocarcinoma group increased on average by 8.93 ± 1.07 ng/ml, and in atypical forms of CL by 2.90 ± 0.23 ng/ml.
The results of the correlative analysis have revealed a significant increase in the content of CA19-9, AFP and |-CG cancer markers in the blood in patients with cervical cancer in 2.2(p < 0.001), in 2.1(p < 0.001) and in 2.5 times(p < 0.001) respectively in comparison with the data of patients with CL.
Moreover, the direct correlation was found between the level of endothelin-1, L-FABP and other oncomarkers: between the level of endothelin-1 and CEA(r = 0,362; p < 0.01), CA125(p = 0.330; p < 0.05), L-FABP(p = 0.466; p < 0.01), | -HCG(p = 0.375, p < 0.01) shows a direct correlation. In addition, a direct correlation was observed between L-FABP and CEA(p = 0.295, p < 0.05), |-CG(p = 0.295; p < 0.05).
As a conclusion, improvement in early detection of CC can be if add to achieved routine of CC diagnostic investigation of CA125, CEA, CA 19-9, AFP and |-CG.
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