ABSTRACT
Due to some drawbacks associated with gene delivery vehicles including viral and non-viral vectors, scientists have continued their efforts to find an ideal gene delivery vehicle. Bacteriophages have been proposed as an attractive alternative gene delivery vehicle in view of their advantages such as protection of transgene, lack of immunogenicity and stability in different conditions. This study has been conducted with the aim of obtaining a construct based on M13 phage and evaluating its capability for delivering transgene to eukaryotic cells. pCMV-Script EX phagemid was constructed by intracellular excision of Lambda-Zap CMV XR vector bearing GFP gene. Packaging of the construct using helper phage resulted in M13-GFP phage particles which used for transfection of human AGS cell line. Finally internalization of the phage particles into the AGS cell line was evaluated by PCR and florescence microscopy. Examination of the treated cells with florescence microscopy indicated that M13-GFP phage particles were able to internalize and express the transgene in eukaryotic cells, but the efficiency of this trasfer was very low. PCR analysis showed that internalization of the M13-GFP gene vehicle to eukaryotic cells was dose dependent. The results indicated that M13 phage could be an appropriate gene delivery vehicle, because it had trivial tropism for eukaryotic cells. This means that after displaying or coupling of appropriate targeting molecules on the surface of phage particles, transgene can effectively be delivered to the target cells
ABSTRACT
Regulation of protein synthesis in the early stage of translation depends on the function of eIF4E factor especially in the form of eukaryotic initiation factor [eIF4F]. Overexpression of eIF4E in multiple cancer types, including malignancies of the prostate, breast, colon, lung, and the hematopoietic system is indicative of the role of this factor in tumorogenesis and promotion of the cancers. In this study we investigated the expression pattern of eIF4E as a new molecular marker in thyroid tumors and their marginal normal tissues. We used semi-quantitative RT-PCR technique to examine the expression of eIF4E in 21 papillary carcinoma tissue specimens and 14 specimens of corresponding marginal normal tissue adjacent to the malignant lesions. Beta 2m gene was considered as an internal control. Rate of expression of eIF4E in different groups were compared with one another by use of SPSS software and data were analyzed by one way ANOVA and t-test. Our data revealed significant expression of eIF4E in all tumor samples compared to non-tumor lesions and normal tissues [P<0.05]. Moreover the expression level was notably increased in malignant tumor samples compared to marginal tissues of the tumors [P<0.05]. The rate of expression was more in tumor samples than non-malignant samples. The results of this study indicated that the rate of expression of eIF4E gene is associated with kind of tumor and grade of malignancy. Also this study confirmed the role of eIF4E gene in tumor progression and development of thyroid tumors. Therefore eIF4E gene expression can be an appropriate indicator for diagnosis of tumors and can be used as a guide for grading of thyroid tumors. This prognostic and diagnostic factor can be considered as a promising therapeutic target for treatment of cancers