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Granulocyte colony-stimulating factor (G-CSF) is the major regulator of hemopoiesis and granulopoiesis. However, overexpression of G-CSF has been implicated in several important processes in tumor biology such as tumor growth, angiogenesis, and metastasis. Four different mRNA isoforms resulting from alternative splicing have been reported for G-CSF (transcript variants 1, 2, 3 and 4). The mRNAs and protein products of splice variants 1 and 2 have been isolated for the first time, from tumor cell lines. In the present study for the first time we isolated the G-CSF transcript variant 4 encoding G-CSF isoform D from a highly malignant tumor cell line (Mehr80) with overexpression of G-CSF. Both the full-length G-CSF isoform B and G-CSF isoform D were cloned from Mehr80 cell line, overexpressed in Escherichia coli as N-terminal glutathione-S-transferase fusion proteins in the form of inclusion bodies and affinity purified by the batch method using glutathione-Sepharose 4B resin. Both fusion proteins were successfully cloned and expressed. Folded recombinant proteins were solubilized from inclusion bodies using sarkosyl, Triton X-114 and CHAPS and purified. The purity of G-CSF isoforms was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and they were clearly detected in western blot analysis using anti-G-CSF polyclonal antibody. The G-CSF plays various roles in physiological and pathological conditions, however to date, the differential function of G-CSF isoforms remains unknown. Considering the fact that G-CSF isoform D was isolated from a highly malignant tumor cell line with overexpression of G-CSF, the role of this splice variant in tumorigenesis requires further investigation. [ABSTRACT FROM AUTHOR]

Objectives This study aimed to assess PD-1gene polymorphism in salivary gland tumors in patients referred to Khalili Hospital in Shiraz. Methods This case-control study evaluated 48 patients with salivary gland tumors and 100 age- and sex-matched healthy controls. First, 5cc blood samples were obtained from patients and transferred to vials containing anticoagulated EDTA. DNA was extracted, and polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) was performed on the samples. The PD-1 gene genotype was determined using the Fermentas kit. After 24 hours of incubation, all the samples were electrophoresed. The genotypes were reported based on the size of bands, and the chi-square test was applied. To compare the alleles, the Fisher's Exact test was applied. The Yates correction was used to compare the genotype and genotypic alleles based on the tumor grade. Results The mean age was 44.81±15.69 years in patients and 46.54± 13.86 years in controls. Statistical analysis did not show any significant difference in PD1 gene polymorphism between the two groups (P=0.098). No significant correlation was found between the genotype frequency and lymph node involvement (P=0.06), tumor genotype (P=0.12), side (right or left) (P=0.22), tumor location (P=0.27), and size or invasion of the tumor to the surrounding tissue (P=0.14). PD1.3 genotype frequency did not differ significantly between malignant and benign tumors (P=0.6). Conclusion This study did not reveal any significant difference in genotype frequency of PD1.3 in the patient and control groups; however, further studies are needed with a larger sample size to obtain more accurate results. [ABSTRACT FROM AUTHOR]

Background: The critical role of interleukin-7 (IL-7) in homeostatic proliferation and T cell survival has made it a promising cytokine for the treatment of various clinical conditions, especially those associated with lymphopenia. Methods: In the present study we expressed recombinant human interleukin-7 (rhIL-7) in Chinese hamster ovary (CHO)-K1 cells. CHO-K1 cells were stably transfected with both circular and linear forms of the pBud-hIL-7 recombinant by electroporation. Expression of rhIL-7 in CHO-K1 cells was confirmed by enzyme-linked immunosorbent assay (ELISA) and dot and western blots. Results: On western blots of transformed cells, a single 25 kDa band was observed, consistent with the expected molecular weight of glycosylated hIL-7. No significant expression difference was observed between cells transfected with circular or linear plasmids. Conclusions: We established a stable CHO-K1 cell line expressing rhIL-7, which we consider to be a promising system for the production of rhIL-7 as a biopharmaceutical. [ABSTRACT FROM AUTHOR]