| ObjectAs it is known, chemotherapy is an important adjuvant therapy for patients with ovarian cancer after operation, for ovarian cancer is still difficult to be discovered at the early stage. However, the cytotox-ic dugs can't selectively kill the cancer cells without any side -effects, the decrease of peripheral blood WBC for myleosuppression caused by cytotoxic dugs contribute to the limitation of regular chemotherapy. Evidence was provided that cisplatin and cyclophosphamide used for ovarian cancer caused apoptosis in several in vivo tumors. For the reasons above, in our study, we detected the apoptosis of total peripheral blood WBC in patients with ovarian cancers before and after chemotherapy. Then analyzed the relationships between the increase of apoptosis and the decrease of peripheral blood WBC, to explore the means to forecast the detection of WBC in patients receiving chemotherapy for ovarian cancers.Subjects and MethodsSubjects;The study included 25 women who received neoadjuvant polyche-motherapy after operation for ovarian cancer. Systematic examination was performed to the patients before the beginning of chemotherapy to be sure that the patients had no immunological disease, infections and others that effect the counts of WBC. Neoadjuvant polychemotherapy consisted of cisplatin and cyclophosphamide The one who received drugs for rapidly declined of WBC was rejected from the study. Methods1. Total white blood cell ( WBC ) counts were performed by certified hematology laboratory.2. Preparation of purified neutrophils: total 2ml of venous blood was collected from patients within 24 hours before chemotherapy and the fifth day after the beginning of chemotherapy using heparin (100u/ ml) as anticoagulant. After incubation at 371 for 2hours in 10-fold volume of PBS ( PH7.4) , erythrocytes were lysed by addition of hypo-tonic lysis. The preparation contained greater than 97% polymorpho-nuclear leukocytes of which 95% were neutrophils cells, Cell viability was greater than 98% as determined by trypan blue exclution.3. Apoptosis analysis(1) Flow cytometry: The percentage of hypodiploid nuclei reflected the relative proportion of apoptosis cells.(2) DNA electrophoresis to detect DNA double - stranded breaks :2% agarose electrphoresis was performed to access DNA fragmentation.4. Statistical analysis; Mean values were compared using the student's two tailed t - test for independent means differences were not regards as significant if p <0.05.Results1 . The change of total WBC and apoptosis before and after chem -otherapy : the counts of WBC are 5.19±1.36±109/Land4. 37± 1.6±109/L the index of apoptosis are4. 01±2. 59%and5. 66± 1. 36%, before and after chemotherapy respectively, both the decrease of WBC and the increase of apoptosis have Statistical significant (p < 0.05) .2. DNA electrophoresis: Agarose gel electrophoresis of DNA were carried out for the detection of DNA double - stranded breaks The presents of DNA ladders demonstrated the breaks of DNA launching the mechanism of apoptosis.3. The correlation between the decrease of WBC and the increase of apoptosis; there is somewhat correlations between them and the coefficient of them is 0. 65.4. The sensitivity of the quality analysis using FCM for clinical early diagnosis is 82% , the speciality is 75% , and the accuracy is 80%.ConclusionThe results demonstrated that the increased apoptosis of WBC induced by cytotoxic drugs contributes to the chemotherapy - associated reduction of leukocyte at some extent. There are somewhat correlations between them. The detection of peripheral leukocyte apoptosis could be helpful for assessing the decrease of WBC. |
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