Purpose: Genetic diseases can be the result of genetic dysfunctions that happen due to some inhibitory and/or environmental risk factors, which are mostly called mutations. One of the most promising treatments for these diseases is correcting the faulty gene. Gene delivery systems are an important issue in improving the gene therapy efficiency. Therefore, the main purpose of this study was modifying graphene oxide nanoparticles by spermine in order to optimize the gene delivery system. Methods: Graphene oxide/APTES was modified by spermine (GOAS) and characterized by FT-IR, DLS, SEM and AFM techniques. Then pEGFP-p53 was loaded on GOAS, transfected into cells and evaluated by fluorescent microscopy and gene expression techniques. Results: FT-IR data approved the GOAS sheet formation. Ninety percent of the particles were less than 56 nm based on DLS analysis. SEM analysis indicated that the sheets were dispersed with no aggregation. AFM results confirmed the dispersed structures with thickness of 1.25± 0.87 nm. STA analysis showed that GOAS started to decompose from 400°C and was very unstable during the heating process. The first weight loss up to 200°C was due to the evaporation of absorbed water, the second one observed in the range of 200– 550°C was assigned to the decomposition of labile oxygen- and nitrogen-containing functional groups, and the third one above 550°C was attributed to the removal of oxygen functionalities. In vitro release of DNA demonstrated the efficient activity of the new synthesized system. Ninety percent of the cells were transfected and showed the GFP under fluorescence microscopy, and TP53 gene was expressed 51-fold in BT-20 cells compared to β-actin as the reference gene. Flow cytometry analysis confirmed the apoptosis of the cells rather than necrosis. Conclusion: It could be concluded that the new synthesized structure could transfer a high amount of the therapeutic agent into cells with best activity.

Background: Breast cancer is one of the most common diseases worldwide that affects women of reproductive age.miR-221andmiR-222are two highly homogeneous microRNAs that play pivotal roles in many cellular processes and regulate the Wnt/β-catenin signaling pathway. Curcumin (CUR), a yellow polyphenolic compound, targets numerous signaling pathways relevant to cancer therapy. The main aim of this study was to compare the ability of chitosan curcumin nanoparticle (CC-CUR) formulation with the curcumin in modulatingmiR-221andmiR-222expression through Wnt/β-catenin signaling pathway in MCF-7, MDA-MB-231 and SK-BR-3 breast cancer cell lines.Method: Chitosan-cyclodextrin-tripolyphosphate containing curcumin nanoparticles (CC-CUR) were prepared. Cytotoxicity of the CUR and CC-CUR was evaluated. Experimental groups including CC-CUR, CUR and negative control were designed. The expression ofmiR-221andmiR-222and Wnt/β-catenin pathway genes was measured.Results: The level ofmiR-221andmiR-222andβ-cateningenes decreased in MCF-7 and MDA-MB-231 cells andWIF1gene increased in all cells in CC-CUR group. However, the results in SK-BR-3 cell line were unexpected; since miRs andWIF1gene expressions were increased following CC-CUR administration andβ-catenindecreased by administration of CUR.Conclusion: Although the composite form of curcumin decreased the expression ofmiR-221andmiR-222in MCF-7 and MDA cells, with significant decreasing ofβ-cateninand increasing ofWIF1gene in almost all three cell lines, we can conclude than this formulation exerts its effect mainly through the Wnt/β-catenin pathway. These preliminary findings may pave the way for the use of curcumin nanoparticles in the treatment of some known cancers.

Schematic representation of experimental design in different groups of mothers and offspring. CTL group: control group; Veh group: vehicle group received normal saline; Ni group: nicotine group received 1 mg/kg/day nicotine, subcutaneously; EtOH group: ethanol group received 3 g/kg/day, 20 % v/v ethanol intraperitoneally; Ni + EtOH group: ethanol plus nicotine which received both.

Purpose: Human butyrylcholinesterase (BChE) serves as a bio scavenger to counteract organophosphate poisoning. It is also a potential drug candidate in several therapeutic fields. Therefore, in the present study, we constructed a new dual-promoter plasmid consisting of Cytomegalovirus (CMV) and human elongation factor 1α (EF-1α) promoters and transfected that into HEK-293 cells using Lipofectamine to enhance the BChE secretion. Methods: The new dual-promoter construction (pBudCE dual BChE) including two copies of the BChE gene was designed and transfected into cells by liposomal structures. The cloned plasmids were evaluated by enzyme digestion and gel electrophoresis analysis. Experimental groups were categorized into the cells transfected by pBudCE dual BChE (treatment), pCMV (positive control) vectors, and nontransfected cells (negative control). BChE gene expression was evaluated by qRT-PCR and the enzyme activity was assessed using modified Ellman’s method. The freeze-thaw process was carried out for analyzing the stability of the pBudCE dual BChE vector. Results: Validation examination of the cloned plasmids confirmed the successful cloning process. The gene expression level and Ellman’s method value in pBudCE dual BChE was higher than the other groups. CMV promoter has also increased the enzyme activity, although the difference was not significant compared with the control group. Interestingly, freeze-thaw cycles followed by several passages did not affect the enzyme activity. Conclusion: The designed construction with CMV and EF-1α promoters could increase BChE gene expression and the activity of the BChE enzyme in HEK-293 cell line. Large-scale production of BChE enzyme can be achieved by using dual-promoter plasmid construction compared to a single-promoter vector to be used in clinical trials.

Context: One of the most common aggressive and primary brain tumors is glioma. The majority of diagnoses are referred to high-grade malignant glioblastoma, which carries the worst prognosis. Still, treatment of brain tumors remains a big challenge for clinicians. This study was designed to investigate the efficacy of gene therapy in the treatment of brain cancer. Methods: Studies use genes as a therapeutic agent in brain cancer treatment even alone or in combination with other treatment methods. Full-text papers, which met the inclusion criteria, were independently assessed by two reviewers. Disagreements were resolved by consultation with a third reviewer. Results: Statistical analysis showed that 50% of the papers used a virus, 36% used polymers, and 14% used cells as carriers to transfect the genes as a therapeutic agent in brain tumor models. Data showed that the estimated size of the brain tumor was reduced by using co-treatment of the gene with one of the conventional therapies. Conclusions: According to the results, co-treatment of the gene with conventional therapies could be more effective than the monotherapy methods.

Breast cancer is one of the most common diseases worldwide. miR-221 and miR-222 are two microRNAs with pivotal roles in many cellular processes which regulate the Wnt/β-catenin pathway. Curcumin (CUR), a yellow polyphenolic compound, targets numerous pathways relevant to cancer therapy. The main aim of this study was to compare the ability of chitosan curcumin nanoparticle (CC-CUR) with the curcumin in modulatingmiR-221andmiR-222expression through Wnt/β-catenin pathway in MCF-7, MDA-MB-231 and SK-BR-3 breast cancer cells. Chitosan-cyclodextrin-tripolyphosphate containing curcumin nanoparticles (CC-CUR) were prepared. Experimental groups including CC-CUR, CUR and negative control were designed. The expression ofmiR-221andmiR-222and Wnt/β-catenin pathway genes was measured. The level ofmiR-221andmiR-222andβ-cateningenes decreased in MCF-7 and MDA-MB-231 cells andWIF1gene increased in all cells in CC-CUR group. However, in SK-BR-3 cells miRs andWIF1gene expressions were increased following CC-CUR administration andβ-catenindecreased by administration of CUR. Significant decreasing ofβ-cateninand increasing ofWIF1gene in almost all three cell lines, indicates that this formulation exerts its effect mainly through the Wnt/β-catenin pathway. These preliminary findings may pave the way for the use of curcumin nanoparticles in the treatment of some known cancers.