Tehran University Medical Journal

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Background: Role of cytokines in regulation of immune system has been the subject of studies and clinical investigations. One of these cytokines, IL-2 has been well initially introduced as T cell Growth factor (TCGF), but subsequently it appeared that IL-2 is one of the important mediators affecting growth, development and activity of T, B, NK and LAK cells. Nowadays this cytokines has extensive use in clinical and research fields of immunotherapy of cancer and infectious disease.

Materials and Methods: In this study, we used Jurcat cell line for production and partial purification of IL-2 106 cell/ml were stimulated by PHA (1 µg/ml) and PMA (10 µg/ml) at the third day of the culture and then supernatant were collected after 22 hrs.

 Results & Conclusion: In order to obtain sufficient amount of IL-2 and eliminate interfacing materials, supernatants were concentrated using Amicon 10 and 30 PM filters. After concentrating, bioassay and Elisa were performed to detect the biological activity and amount of produced IL-2. Reversed phase-HPLC was used to confirm the IL-2 identity and purification.

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Background: Ewing's sarcoma is one of the most malignant tumors in children and young adults. Only a few established cell lines of Ewing's sarcoma have been reported, which makes it difficult to study the biological features of these tumors. We have recently established a new Ewing's sarcoma cell line designated SS-ES-1, originating from a thoracic tumor of a 16-year-old female patient. The SS-ES-1 cells have been grown continuously in culture for over 90 passages. In this report, some characteristics of SS-ES-1 cells are presented.
Methods: The cells were grown in DMEM medium supplemented with 10% fetal bovine serum (FBS), 100 mg/ml streptomycin and 100 U/ml penicillin in a humidified atmosphere with 7% CO2 at 37 ºC. The cells were immunocytochemically characterized using a panel of monoclonal and polyclonal antibodies. Furthermore, the chemo-sensitivity of this cell line to some anticancer drugs was assessed using MTT assay and IC50 values were determined.
Results: Morphologically, the SS-ES-1 cell line is composed of poorly differentiated small round cells growing in a multilayer pattern. The immunocytochemical staining demonstrates strong reactivity to CD99, cytokeratin, neurofilament, p53 and Ki67 proteins, but no reactivity to GFAP. Based on IC50 values, SS-ES-1 cells display considerable sensitivity to vinblastine (2±0.7 pM), followed by vincristine (0.3±0.12 nM), doxorubicin (0.05±0.03 µM), etoposide (0.64±0.28 µM) and cisplatin (0.67±0.45 µM).
Conclusions: In conclusion, the SS-ES-1 cell line demonstrates unique cellular properties, which make it a useful model for studying various aspects of the biology of Ewing's sarcoma.

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Background: Vascular endothelial growth factor (VEGF) has mitogenic effect for endothelial cells and is an important mediator of tumor expansion, metastasis and angiogenesis in vivo. Isosorbide dinitrate, as a nitric oxide donor, has been widely used in treatment of many cardiovascular diseases such as congestive heart failure and acute coronary syndromes. Furthermore this drug was found to have inhibitory effect on angiogenesis, tumor growth and metastasis in vivo. In the present study we evaluated the isosorbide effect on the VEGF production using some human leukemic cell lines.

Methods: Human leukemic MOLT-4, JURKAT and U937 cells were cultured in complete RPMI medium. The cells at the exponential growth phase were then incubated with different concentrations of Isosorbide (4´10^-7 -4´10^-4 M) in the presence or absence of PMA (25ng/ml) for 24 hours. The VEGF concentrations in the culture supernatants were measured by enzyme immunoassay kits (R&D systems) according to the manufacturer's instructions.

Results: The level of VEGF produced by the human leukemic cell lines which was treated with different concentrations of isosorbide, did not show any significant difference with untreated control cells.

Conclusions: The results of this study showed that isosorbide had no significant effect on VEGF production. Our findings suggest that anti-angiogenesis effect of isosorbide could be mediated through VEGF-independent mechanism(s). Further studies are warranted to determine definite isosorbide effect on VEGF and other angiogenic factors production in patients as well as animal models.

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Background: Lactobacillus species are genetically diverse groups of Lactic Acid Bacteria (LAB) that have been introduced as probiotics, because of some characteristics such as their anti-tumor properties, helping the intestinal flora balance, production of antibiotics, stimulation of host immune response, etc. The aim of this study was to investigate the effects of cytoplasmic extraction and cell wall of Lactobacillus species isolated from the intestine of common carp on human chronic myelocytic leukemia or K562 cancer cell lines.
Methods: The intestinal contents of 115 common carp captured from the natural resources of West Azerbaijan province in Iran were examined for LAB. After isolation, the identification of Lactobacilli was done according to traditional and molecular bacteriological tests. Subsequently, a suspension of each bacterium was prepared and the protein content of the cytoplasm was extracted. Cell wall disintegration was done by cell lysis buffer and sonication. The effects of cytoplasmic extraction and cell wall on K562 cell line proliferation were investigated by MTT assays.
Results: The cytoplasmic extraction of the isolated Lactobacilli had significant (p<0.05) anti-proliferative effects on K562 cells. The cytoplasmic extractions of Lactobacillus paracasei and Lactobacillus casei inhibited K562 cell proliferation by 66.56% and 54.28% at 83.33 μg/ml concentration, respectively.Nevertheless, the Lactobacillus cell wall could not inhibit the proliferations of K562 cells (p<0.05).
Conclusion: In this study, the cytoplasmic extractions of the isolated Lactobacilli from the intestine of common carp had anti-proliferative effects on K562cell line.