Relation between Serum cyclo oxygenase-2 values and Tumor characteristics in breast cancer patients
Fatima Moselmani , Jumana Al- Saleh
Pharmacy College- Damascus University- Damascus- Syria.
*Corresponding Author E-mail: fatimamoslmani.1990@gmail.com
ABSTRACT:
Background: Breast cancer hasa high incidence and death rates, and cyclooxygenase-2 (COX-2) is a promising enzyme in as prognostic and monitoring factor. Objective: Determination the relation between serum COX-2 concentrationand the stage and grade of breast cancer as a prognostic parameters. Methodology: This study is prospective, cross sectional type, include 30 women with breast cancer, met the study criteria. the serum COX-2 value was calibrated. The data was entered into the computer's SPSS program. Results: Patients were 48.33 ± 7.84 years old, and 70% of tumors in the first and second stages, while more than half of the tumors were in the third grade. The mean of serum COX-2 values was 372.31 ± 173.42 U/L.Serum COX-2 level was positively and significantly correlated with tumor stage and grade(P <0.01). Conclusion: Serum COX-2 values correlate with the stage and grade in breast cancers.
KEYWORDS: Breast Cancer, Cox-2.
INTRODUCTION:
Breast cancer is a global concern because of its high and rising incidence, and is one of the leading causes of cancer deaths in the world[1].
Cyclooxygenase (COX) interferes with the formation of biological media starting with arachidonic acid, synthesizes prostaglandins and intervenes in inflammatory events. COX-2 is an inducible type of COX, undetectable in many normal tissues, becoming abundant in large activated phagocytes and other cells at inflammation sites[2,3].
The expression is excessive in many tumor tissue samples (lungs, esophagus, breast, colon, rectum, pancreas)[2-6]. The expression of COX-2 can interfere with tumor genesis, angiogenesis, inhibition of apoptosis, modulating the immune response, and also contribute to tumor invasion, and metastasis[7-9].
Breast cancer cells have an overexpression of the COX-2 gene. Meta-analysis found an overexpression of the COX-2 in 27.9% -81.4% of cases of breast cancer[3]. COX-2 interferes with the pathogenesis of breast cancer by interfering with many breast cancer-related processes through activating signal pathways of PKC, PKA or TGFβ via specific receptors for PGE2[10]. Also COX-2 and its products affect tumor cell growth by affecting the DNA synthesis[11,12].
Excess expression of COX-2 is associated with poor prognostic variables such as lymph node metastases, poor cell differentiation, and large tumor size[13,14]. A meta-analysis recognized that the high expression of COX-2 was significantly associated with a short 5-years survival rate and a decrease in tumor-free duration[3]. A study found a relationship between a high expression of COX2 in tumor cells and invasive and metastatic breast cancers[15].
Previous studies focused on the histological specimens, which made the results remain limited in the research field, not for practical use. Therefore, this study relied on the manufacturer's data for COX-2 kits on the calibration of the concentration of COX-2 in the serum of the patients to determine its concentration, and its association with tumor characteristics.
METHODS:
The study is a prospective, cross sectional study, conducted in the laboratories of the Faculty of Pharmacy, and University Hospitals- Damascus University- Syria, during the period 2017-2018. It included 30 women with breast cancer. Every woman gave her informed consent before entering the study.
The women underwent surgery to remove the tumor mass according to the evaluation of surgeons, and the histological sample was sent to the pathology. Accordingly, the stage and grade of the tumor were determined.
Exclusion criteria:
Women with two-sided breast cancer, a current or previous tumor story, or chemotherapy or radio-therapy were excluded. Also We excluded women with known diseases affecting serum COX-2 values such as acute infections or immune diseases.
COX-2 assay:
Concentrations of COX-2 in the serum were determined using a COX-2 ELISA kit,as described in the protocol provided by the manufacturer(SunRed Biotechnology Company, Shanghai, China):
Coagulation the specimen at room temperature 10-20 mins, centrifugation 20-min at the speed of 2000-3000 rpm, remove supernatant. If precipitation appeared; centrifugal again.
Added to monoclonal antibody enzyme well which is pre-coated with Human COX-2 monoclonal antibody, incubation; then, added COX-2 antibodies labeled with biotin, and combined with Streptavidin-HRP to form immune complex; then carried out incubation and washed again. Then added Chromogen Solution A, B, gently mixed, incubated for 10 min at 37 C away from light. The reaction wasterminated by addition of stop solution. The optical densit (OD) was measured within 15 min after adding the stop solution, at 450nm wavelength in an EnzymeReader.
A standard curve was constructed using the standards' concentration provided in the kit, and the OD values of the sample applied on it to calculate the corresponding sample's concentration.
Data analysis:
Data were entered into the computer and analyzed using SPSS-17 (SPSS, Inc., Chicago, IL). The data were presented in descriptive form as percentage, mean±standard deviation. The analysis was performed on the t-student test with the variable considered statistically significant when the value of the significance level is less than 0.01.
RESULTS:
The study involved 30 women with breast cancer. The median age of patients was 48.33 ± 7.84 years (range: 33-63 years), and Table 1 shows patients characteristics.
Table (1): patient's characteristics
|
characteristics |
Results |
|
Age |
48.33±7.84 (range: 33-63 y) |
|
Tumor type |
|
|
ductal |
30 (100%) |
|
Tumor stage |
|
|
1 |
5 (16.7%) |
|
2 |
16 (53.3%) |
|
3 |
6 (20%) |
|
4 |
3 (10%) |
|
Tumor grade |
|
|
1 |
2 (6.67%) |
|
2 |
12 (40%) |
|
3 |
16 (53.3%) |
The study recorded a mean of serum COX-2 values of 372.31 ± 173.42 U/L. The correlation of COX-2 values with tumor characteristics was studied. Table 2 shows the results.
Table (2) COX-2 values
|
|
COX-2 valus |
p-value |
|
stage |
|
|
|
1 |
261.8±65.2 (207.3-316.3) |
0.00 |
|
2 |
310.4±191.9 (181.4-439.3) |
|
|
3 |
507.5±116.0 (418.3-596.6) |
|
|
4 |
546.7±32.6 (254.1-839.4) |
|
|
grade |
|
|
|
1 |
189.4±75.3 (110.4-268.4) |
0.00 |
|
2 |
244.8±74.7 (187.3-302.2) |
|
|
3 |
522.0±90.9 (471.6-572.4) |
COX-2: Mean±SD (95% CI)
DISCUSSION:
The relationship between COX-2 expression and tumors has become a promising point in oncology researches. COX-2 plays an important role in the initiation of breast cancer.[2,3].
The study found a significant positive correlation of serum COX-2 level with tumor stage and grade. Such correlation has been recorded by previous reports for histological expression of COX-2 in breast cancer cells. A meta-analysis of studies found that increased COX-2 expression was significantly associated with large tumor size, positive lymph nodes,[3] and other studies found That histological expression of COX-2 is associated with metastases of axillary lymph nodes, poor cell differentiation, and large tumor size[13.14].
This study may be the first study looking for the importance of the titer of the value of COX-2 in serum in breast cancer patients, with previous studies focus on the positive tissue expression of COX-2, hence the importance of this study because of the difference in the ease of serological calibration and low cost, and acceptance of it, also allows for the use of repeated assays of COX-2 in monitoring therapeutic response and subsequent tumor recurrence.
The study recommends a prospective study to monitor changes in serum COX-2 values during and after treatment of women with breast cancer.
CONFLICT OF INTEREST:
The author declares no conflict of interest.
ACKNOWLEDGEMENT:
I would like to thank Dr. Gomanah Al-Saleh for her advices.
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Received on 29.10.2019 Modified on 19.12.2019
Accepted on 12.02.2020 © RJPT All right reserved
Research J. Pharm. and Tech 2020; 13(9):4320-4322.
DOI: 10.5958/0974-360X.2020.00763.5