Modares Journal of Biotechnology

Abstract Aims: Programmed cell death is a vital cellular process that is highly conserved in evolution. Apoptosis, as a common mode of programmed cell death, is disturbed in the most human malignancies and leads the resistance of cancers to current treatment strategies. Caspase 9 is a key protein in mitochondrial apoptosis. Activated Caspase 9 leads to activation of Caspase 3/7, initiating a caspase cascade and killing cell. In this study, Caspase 9 gene was cloned into pcDNA3.1(+) and its expression and function evaluated in cell.
Methods: PCR amplification of Caspase 9 was performed by specific primers and ligated into pcDNA3.1(+) after double-digestion with KpnI and BamHI. After sequencing, pcDNA/Caspase 9 was transfected into SH-SY5Y cells and treated with doxorubicin. Caspase 9 function was determined by its effect on cell death level by trypan blue and PI staining, and Caspase 3 activity, and its expression in cells measured by western blotting.
Finding: Caspase 9 gene cloning was done and its expression in cell defined by western blot. Overexpression of Caspase 9 led to autoprocessing following homodimerization and induction of cell death and also increased cell sensitivity to doxorubicin treatment and declined cell viability.
Conclusion: The cloned Caspase 9 was functional in cell and enhanced apoptosis in the treated cells by doxorubicin through self-activation and subsequently amplification of Caspase 3 activation.

Abstract Chemotherapy is one of the methods used in the treatment of cancer. Not being targeted has many side effects for the patient. The use of nano-carriers, including nano-liposomes, is a very effective way to target chemotherapy. In this study, a liposomal system containing doxorubicin was designed and evaluated for its effect on lung cancer cells.
In this in vitro study, two liposomal systems were prepared by the method of thin-film hydration and using different concentrations of phosphatidylcholine and cholesterol. Doxorubicin was then loaded into the systems. Finally, one of the systems was selected based on the drug loading rate and drug release pattern. At the end, the selected drug delivery system in terms of particle size, zeta potential, liposomal vesicle appearance, and the interaction between the drug and the system was investigated.
The selected liposomal system contains doxorubicin with an encapsulation efficiency of 58.89%, size of 273 nm, dispersion index of 0.458 and zeta potential of -35.7 mv. Doxorubicin release from liposomes was controlled and no chemical interaction was observed between liposomes and referees. Liposomal vesicles are also spherical and have a smooth surface.
The results of this study show that nano-liposomes can be prepared with appropriate formulation containing doxorubicin using nanotechnology. Which has good physicochemical properties. Therefore, this liposomal system can be recommended for further cancer-related research.

Abstract Chemotherapy is one of the most effective and common treatments for cancer. Multi-drug resistance and drug side effects are one of the major obstacles to successful chemotherapy. To address these limitations and achieve better drug efficacy, nanosystem-based combination therapy offers a promising approach. This study aimed to synthesize, characterize, and investigate the synergistic effect of nanoliposomes loaded with doxorubicin and epigallocatechin-3-gallate (EGCG) on MCF-7 breast cancer cell lines. In the present study, nanoliposomes were prepared by passive loading and thin-film hydration. The characterization of nanoliposomes such as size distribution, zeta potential, the loading rate, drug release profile, and toxicity were measured. The mean diameter of nanoliposomes was 82.5 nm, their surface charge was -24.2 mV and drug loading was about 80%. The interaction of doxorubicin and EGCG with nanoliposomes was mediated by electrostatic and van der Waals bonds and EGCG has a deceasing effect on the doxorubicin release profile but the observed differences are not significant. The toxicity and viability data indicate that the simultaneous use of these two drugs increased the toxicity of the cells. Nanoliposomes containing doxorubicin were not able to reduce viability to below 50% in monotherapy with 5×10^-3 μM. While, the amount of viability dramatically reduced to below 50%, in combination with EGCG, resulting as combination chemotherapy. Consequently, the concomitant administration of EGCG with doxorubicin may be a suitable candidate for chemotherapy.

Abstract Iron oxide nanoparticles are one of the nano carriers that are suitable for novel drug delivery systems due to low toxicity, biocompatibility, loading capacity and controlled drug delivery to cancer cells. The purpose of this study is the synthesis of coated iron oxide nanoparticles for delivery of Doxorubicin (DOX) and its effects on cancer cells.
In this study, Fe₃O₄ magnetic nanoparticles were synthesized by Polyol method, and then doxorubicin was loaded onto PEGylated iron oxide nanoparticles. FT-IR was used to ensure PEG binding to nanoparticles and loading the drug onto nanoshell. Comparison of the mean size and the crystalline structure of nanoparticles were performed by TEM and X-ray diffraction pattern. Then, the effect of cytotoxicity was evaluated on AGS and MCF-7 cancer cells by MTT assay.
According to FT-IR results, the presence of O-H and C-H bands at 2927 cm^-1 and 3392 cm^-1 peaks correlated with PEG binding to nanoparticles. XRD pattern showed the cubic spinel structure of trapped magnetite nanoparticles carrying medium with a mean size of 14 nm. 21.67% of Doxorubicin was loaded into Fe₃O₄-PEG nanoparticles, which the highest drug release recorded during the first 24 hours. MTT assay at 24, 48 and 72 h treatments showed that with increasing concentrations of doxorubicin loaded Fe₃O₄-PEG nanoparticles from 0 to 50 μm, the cytotoxic effects of the drug gradually increase.
This study showed that PEGylation of iron oxide nanoparticles and using them in drug delivery process to increase the effect of Doxorubicin on AGS and MCF-7 cancer cells