Disseminated tumor cells and enhanced level of some cytokines in bone marrow and peripheral blood of breast cancer patients as predictive factors of tumor progression.

BACKGROUND: In recent years, the presence of disseminated tumor cells (DTC) in bone marrow (BM) of patients with breast cancer (BC) is considered as an important clinical feature of the distribution process. However, relapse often occurs in spite of the negative results of the bone marrow cytology. This suggests the need to find additional signs of the possibility for predict the recurrence. AIM: to detect DTC in BM and determinate cytokine status of peripheral blood (PB) and BM of primary BC patients for prognosis of tumor recurrence. PATIENTS AND METHODS: 72 BC patients with histologically proven diagnosis were enrolled into study. 31 patients with progression of disease and 41 patients with clinical stabilization (conditional remission) were included to "progression" and "remission" group respectively. This division of BC patients was conditional and was made during the 3 years study. The presence of DTC in BM was detected by immunocytochemical analysis. Plasma levels of TNF-α, M-CSF, IFN-α were defined by bioassay tests. Plasma levels of IL-6, TGF-ß1 and VEGF were determined by ELISA. BM and PB BC patients were obtained before treatment. RESULTS: In our study DTC in samples of BM were detected in 50% BC patients of progression group. It was found that most significant addition markers of tumor progression with presence of DTC in BM are the levels of cytokines such as TNF in BM and PB, CSF-1 and IL-6 in PB and endogenous IFN in BM and PB of BC patients. In patients of disease "progression" group the levels of TNF in BM were increased by 45.8% (p < 0.01), the levels of CSF-1 and IL-6 in PB were increased more than by 70-80% (p < 0.05). CONCLUSION: Comprehensive detection DTC in BM and identification the level of TNF, IFN, CSF-1, IL-6 in PB and BM of BC patients could be one of the ways for prognosis the metastatic process and correction antitumor individualized therapy.

Karyotype alterations in human lung adenocarcinoma cells after long-term action of interferon-alpha.

AIM: To estimate the effect of long-term IFN treatment of human non-small-cell lung cancer cell line A-549 on their karyotype characteristics and on the clonal structure of cell population. METHODS: Cytogenetic research was performed by standard methods using routine and differential staining. Cytogenetic characteristics were estimated per 1000 cells (ppm, (per thousand)). RESULTS: Cytogenetic analysis of IFN-modified A-549 human lung cancer cells had demonstrated far-going changes in their population structure. It was shown that long term cultivation with IFN altered the chromosome modal class of A-549 cells, induced the domination of hromosomes with certain molecular markers: the number of metaphases with der (6) t (6; 1) chromosomal rearrangement increased significantly (from 6% to 80%, p CONCLUSION: Long-term IFN effect results in alterations of cytogenetic properties of A-549 human lung cancer cells.

Expression of drug resistance proteins in triple-receptor-negative tumors as the basis of individualized therapy of the breast cancer patients.

AIM: To evaluate the efficacy of the application of various chemotherapy schemes based on the immunohistochemical study of expression patterns of proteins associated with the drug resistance P-glycoprotein (P-gp), glutathione-S-transferase (GST), metallothioneins (MT) of breast cancer (BC) patients with the triple-receptor-negative (RE(-), RP(-), HER-2/neu(-)) cancer. METHODS AND RESULTS: P-gp, GST and MT expression in BC-biopsy samples from 60 BC patients was evaluated by immunohistochemical analysis. The results of the clinical observations showed that 3-years relapse-free survival rate of the patients of with P-gp, GST and MT-positive tumors treated with taxoter + adriablastin / taxoter + cyclophosphamide (TA/TC), gemcitabine + carboplatin, or TC + bevacizumab was 61.5%, 78.6% and 81.2% respectively, vs 41.2% in the control group with P-gp, GST and MT-negative tumors treated with adriablastin + cyclophosphamide (p<0.05), while overrall suvival rates were 84.4%, 92.6% and 93.8% respectively vs 70.6% in the control group (p<0.05). CONCLUSION: The study points on the possibility to elevate the efficiency of polychemotherapy by its individualization based on the expression patterns of P-gp, GST and MT on tumor cells of the patients with the triple-receptor-negative BC.

Modifying influence of prolonged action of interferon on phenotypic characteristics of human lung cancer cells in vitro.

AIM: To study modifying influence of interferon (IFN) on some phenotypic properties of human non-small-cell lung cancer cells (NSCLC) upon prolonged exposition of the cells with IFN. MATERIALS AND METHODS: A-549 cells were cultivated with IFN at increasing concentrations for a long period of time (up to 1 year). Cell morphology and ultrastructure were studied by light and electron microscopy. Expression of adhesion protein E-cadherin, and vimentin, cytosceleton protein associated with tumor cell migration and invasion, antigen of proliferating cells Ki-67, angiogenesis-stimulating protein VEGF were studied using the method of immunocytochemistry. Autonomity of the cell growth was studied with the use of colony formation in soft agar, platting efficiency assay, and growth in serum-free medium. RESULTS: It has been shown that prolonged action of IFN results in significant and irreversible inhibition of manifestation of malignant phenotype: decreased of proliferative potential and inhibited autonomy of the cells; in complicated cell ultrastructure; in decreased expression vimentin and in increased expression of E-cadherin. Also, an inhibiting influence of IFN on expression of EGF receptors and VEGF in tumor cells have been shown. CONCLUSIONS: The data are showing that prolonged exposition of NSCLC cells to IFN is accompanied by stable phenotypic alterations of the cells directed on significant loss of malignancy and their shift to more differentiated phenotype.

Effect of anticancer drugs on production of transforming growth factor and expression of p53 AND Bcl-2 proteins by MCF-7 and T47D cell lines of human breast carcinoma.

AIM: To compare the capability of methotrexate, cisplatin, doxorubicine and vincristine to induce production of the transforming growth factor beta(1) (TGF-beta(1)) in two cell lines - MCF-7 and T47D - of human breast carcinoma, as well as to study sensitivity of these cells to TGF-beta(1) and mentioned anticancer drugs. MATERIALS AND METHODS: ELISA for detection of TGF-beta content in conditioned culture media and Western-blot analysis of the proapoptotic p53 and antiapoptotic Bcl-2 proteins were applied. RESULTS: T47D cells showing higher resistance to growth inhibiting effect of TGF-beta(1) were also refractory to cisplatin. There was no difference between MCF-7 and T47D cells in their sensitivity to methotrexate and doxorubicine, although T47D cells were more sensitive to vincristine. It was found that methotrexate and vincristine did not affect TGF-beta(1) production, while doxorubicine used at a dose of 1-100 ug/ml, significantly induced TGF-beta(1) production in both cell lines. p53 expression in T47D cells was higher than in MCF-7 cells where only doxorubicin induced strongly p53 expression. It should be noted, that Bcl-2 was better expressed in MCF-7 cells, while it was almost undetectable in T47D cells. CONCLUSION: In cells of human mammary carcinoma of MCF-7 and T47D lines doxorubicine, unlike vincristine and methotrexate, in dose depending manner induces production of TGF-beta(1). TGF-beta(1) production in carcinoma cells was associated with doxorubicine-mediated p53 expression in MCF-7 cells or high basal level of p53 in T47D cells. The cells of MCF-7 line were more sensitive to growth inhibition by exogenous TGF-beta(1) and to cisplatine action than T47D cells, but there was no difference between these cell lines in sensitivity to other anticancer drugs.

[The evaluation of the efficacy of adjuvant laferon therapy in the combined treatment of patients with ovarian cancer based on the monitoring of the tumor marker Ca-125 in the blood serum]. / Otsinka efektyvnosti ad'iuvantnoï laferonoterapiï u kompleksnomu likuvanni khvorykh na rak iaiechnyka na pidstavi monitorynhu pukhlynnoho markera Ca-125 v syrovattsi krovi.

Relationship was established of blood level of CA-125 in patients with ovarian carcinoma (OC) to histologic form of tumour, with the highest CA-125 levels being recordable in patients with poorly differentiated OC, lower ones in those having serous form of OC, and the lowest in patients with mucinous form. Complex treatment schemes involving laferon make for striking and stable reduction of blood levels of CA-125 in patients presenting with serious and mucinous forms of OC and, strictly speaking, do not affect this same parameter in patients with poorly differentiated OC.

[The role of interferon in anti-tumor resistance]. / Rol' interferon v protivoopukholevoi rezistentnosti.

The author's own data and those available in literature on the experimental development of the interferon participation in anticarcinogenesis are reviewed. The mechanisms of antitumoural effects of exogenic interferon are analyzed and expediency of its incorporation into the complex antitumoural therapy aimed at prevention of the metastatic development is substantiated. The data are presented on the individual sensitivity of the human mammary gland cancer (the primary tumour and metastases) to interferon.

[The cellular sensitivity of the primary tumor and its metastases in breast cancer patients to cytostatics, interferon and tumor necrosis factor]. / Issledovanie chuvstvitel'nosti kletok pervichnoi opukholi i metastazov u bol'nykh rakom molochnoi zhelezy k tsitostatikam, interferonu i faktoru nekroza opukholei.

Samples of tumor tissue obtained from 47 patients with primary and metastatic breast cancer were implanted under the renal capsule of mice (SRCA-test) to assess individual sensitivity of these malignancies to cytostatics, recombinant interferon (rIFN) and--in some cases--to tumor necrosis factor (TNF). Primary tumor was shown to respond to cytostatics and rIFN in as few as 33.3 and 26.7% of cases, respectively. Xenografts of metastases displayed higher rates of response to all the drugs studied, viz. 64.2, 86.7 and 90% for cytostatics, rIFN and TNF, respectively.

[Normalizing effect of interferon on the system of mixed-function oxidases in the liver of mice with Lewis lung carcinoma]. / Normaliziruiushchee deistvie interferona na sistemu mnogotselevykh oksidaz pecheni myshei s kartsinomoi legkogo L'iuis.

The effect of interferon (IF) on the activity of multipurpose hepatic oxidases (MHO) in animals in the process of metastatic tumour development as well as interaction of IF and vinblastine (VBL) in the cell culture have been studied. IF activates MHO under conditions of their inhibition at the beginning of the tumour development and inhibits them to the normal level under activation in the postmetastatic period. Contrary to the data obtained in vivo IF does not decrease the toxicity of VBL for the tumour cells in vitro. The data obtained evidence for the modulating effect of IF on the MHO system under the tumour development.

[Influence of interferon on the toxic effect and antitumor activity of vinblastine in the combined therapy of metastatic Lewis carcinoma]. / Vliianie interferona na toksicheskii éffekt i protivoopukholevuiu aktivnost' vinblastina pri kombinirovannoi terapii metastaziruiushchei kartsinomy L'iuis.

Antitumour and antimetastatic effects of the combination of vinblastine (VLB) and L-cell interferon were examined. It is shown that mice with 3LL carcinoma are more sensitive to common toxic effect of VLB than intact animals. Interferon (IF) decreases the toxic effect of VLB in tumour-bearing mice, and thus provides a possibility to increase the total therapeutic dose of the cytostatic. The combination of VLB and IF showed a synergistic inhibitory effect on the development of carcinoma metastases and significantly increases the survival rate of the tumour-bearing mice. Neither synergistic nor additive antitumour effects of the combined therapy were observed on the primary tumour. IF suppresses the primary tumour growth more significantly when given by intratumoural injection. At the same time the inhibition of metastase development was more efficient with intraperitoneal injection of IF.

[Interferon-forming properties of the leukocytes in assessing the functional activity of the regional lymph nodes in cervical cancer]. / Interferonoobrazuiushchie svoistva leikotsitov v otsenke funktsional'noi aktivnosti regionarnykh limfaticheskikh uzlov pri rake sheiki matki.

In the present work the authors have employed their personal modification of the technic of interferon reaction of lymphocytes, which consists in using the lymphocytes isolated from regional lymph nodes but not from the peripheral blood as described in the literature. The titres of interferon produced in vitro by the lymphocytes of lymph nodes, removed during the operation in cervical cancer patients, were shown to correlate well with the clinico-morphological changes, indicating the rate of the regional lymphoid tissue response. The mentioned correlation speaks in favour of the prognostic value of the interferon reaction of regional lymph nodes lymphocytes in cervical cancer.

[Tilorone, a synthetic immunostimulator possessing antitumor activity]. / Tiloron--sinteticheskii immunostimuliator, obladaiushchii protivoopukholevoi aktivnost'iu.

The authors report the results of studies on antitumor, immunostimulating and interieron-inducing properties of diethylamine fluorene derivative--tiloron, synthesized at Odessa laboratories of IGIC of the Ukrainian SSR Academy of Sciences by the originally elaborated method. It was found that tiloron does not suppress leucopoiesis, stimulates a number of protective reactions of the organism, including the factors of humoral and cell immunity, especially manifested under immunosuppression. Tiloron is shown to possess the definite antitumor activity being retained in peroral administration of the drug. Tiloron used together with cytostatic agents results in enhanced suppression of experimental tumors growth. Antitumor and immunostimulating action of tiloron on the experimental patterns under study is believed by the authors to be somewhat related with its interieron-inducing ability.