فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 53 (زمستان 1402)

Farnesol and gingerol are two powerful plant active ingredients that have proven anti-cancer and antioxidant effects. In this study, the expression changes of Caspase 8, Matrix metalloproteinases 2, Matrix metalloproteinases 9 and Vascular endothelial growth factor genes were studied in MCF-7 and SKBR3 cancer cell lines treated with niosomes containing farnesol and gingerol.

MCF-7 and SKBR3 cells were treated with niosome containing farnesol and gingerol, combination of two drugs, single drug niosome formulation and free drug for 48 hours. Gene expression changes were investigated using Real-Time PCR technique and statistical analysis was done at 0.05 level with GraphPad Prism software.

Niosome codelivery containing two drugs (N-Far/Gin) caused a significant increase in the level of caspase 8 gene expression and also a significant decrease in the regulation of the expression of Matrix metalloproteinases 2, Matrix metalloproteinases 9, and Vascular endothelial growth factor genes (P<0/001). It had a higher significance level than the treatment with other formulations.

It seems that farnesol and gingerol-loaded niosomes have great anti-apoptotic, anti-metastatic, and anti-angiogenic potential in the prevention of breast cancer. Further in vivo experiments and clinical studies can elucidate the potential of N-Far/Gin to be applied as a medicinal agent in breast cancer treatment.

Oncolytic viral therapy is known as promising approach for cancer treatment. Oncolytic viruses like adenoviruses can proliferate in cancerous cell and disintegrated these cells.  Serotype 5 adenovirus attached on surface of host cell via coxsackie-adenovirus receptors. The expression rate of this receptor is highly variable in surface of cancerous cells. The aim of the present study was production of an oncolytic adenoviruses with high selectivity feature for gastric adenocarcinoma cells (AGS) that can proliferate in this cell and destroy them.

In this experimental study, the octreotate sequence was cloned into HI-loop of the adenovirus fiber. Thus, the pAd5TATE (wt) and pAd5TATE (ΔE1, ΔE3) recombinant vectors were constructed. pAdZ-TATE (wt) vector was synthesized using pADZ cloning system. Ad5TATE (wt) and Ad5TATE (ΔE1, ΔE3) recombinant viral particles were synthesized and propagated in HEK293 cells. Using the calcium phosphate procedure, the HEK293 cell line was transfected with the recombinant vectors and the new virus particles were produced. The AGS cell line was transfected by the Ad5 (wt) and Ad5TATE (wt) and the cell destruction of these cells were investigated in different times. Using the MTT test the cell surveillance rate was evaluated in AGS cell line that treated with recombinant vector. Cell destruction effects (CPE) started in less than 18 hours and were completed 72 hours after infection.

The titer of recombinant viruses was calculated by the TCID50 method as 107 (for 100 Lµ of viral solution). Also, Ad5TATE (ΔE1, ΔE3) virus titer was 107 TCID50/ml by the TCID50 method. The results showed that Ad5TATE (wt) could infect AGS cells compared to Ad5(wt). The results of the MTT test showed that with the increase in the concentration of viral particles and storage time, the number of live AGS cells decreased significantly.

The findings of this research showed that the Ad5TATE (wt) recombinant virus could specifically infect gastric adenocarcinoma (AGS) cancer cells and destroy them after replication. Therefore, the Ad5TATE (wt) virus can be used as an oncolytic agent against gastric adenocarcinoma (AGS) cancer cells.

Breast cancer is the most common type of cancer among women, which occurs in the epithelial tissue of the mammary gland. Various environmental, genetic and epigenetic factors play a role in the occurrence of breast cancer. According to the previous studies, one of the most important epigenetic changes involved in the occurrence of breast cancer is the dysregulationof the expression level of microRNAs. Therefore, the aim of this study was to investigate the expression level of miR-509-3-5p and miR-596 in breast cancer tumor tissue.

In the bioinformatics analysis, the datasets related to miRNAs (GSE40525 and GSE45666) were prepared from the GEO database. Data analysis was performed using the Affy package in R software. Sampling was done from 100 women with breast cancer and 100 control. Tissue RNA was extracted using Trizol solution. Then, using the cDNA synthesis kit, DNA was synthesized from the extracted RNAs, and the expression level of miR-509-3-5p and miR-596 was investigated using Realtime PCR method.

According to the bioinformatics results, the expression level of miR-509-3-5p and miR-596 in breast cancer tissue reduced compared to healthy tissue. The expression level of miR-509-3-5p and miR-596 in tumor tissue was significantly lower than normal tissue (p<0.05).So these results confirmed the results of bioinformatics studies.

According to the results of the present study, which showed a decrease in the expression of miR-509-3-5p and miR-596 in breast cancer, it can be said that these miRNAs can be used as important diagnostic and therapeutic biomarkers in breast cancer.

Histones are replaced by protamines to package sperm head DNA during mammalian spermatogenesis. Protamine genes variation cause sperm DNA damage and is affect infertility in men. Therefore, this study aim was investigation on association of two rs737008 in PRM1 gene and rs4780356 in PRM2 gene polymorphisms with azoospermia and oligospermia in Iranian idiopathic infertile men.

DNA extraction from blood samples of 96 idiopathic infertile men with azoospermia and oligospermia and 100 normal control men. Prevalance of two studied polymorphisms was identified by restriction fragment length polymorphism (PCR-RFLP). Then results were confirmed with sequencing.

A mutant allele frequency of rs737008 in PRM1 gene polymorphism was difference in patient and control groups but statistical analysis showed no significant association between this polymorphism prevalance among case and control groups (P>0.05). By genotyping shown no existent rs4780356 in PRM2 gene polymorphism in anyone of case and control groups.

This study finding indicated there was not association with prevalence of two rs737008 in PRM1 gene and rs4780356 in PRM2 gene polymorphisms with oligospermia and azospermia and idiopatic male infertility in Iranian population. More studies need to demonstrate the role of these two polymorphisms with idiopathic infertility in Iranian men.

COVID-19, an infectious viral disease caused by severe acute respiratory syndrome (SARS-CoV-2) with more than 260 million infections as of December 2019, is a serious threat to the health and economy of human societies. Designing and producing a suitable vaccine can Reduce the incidence of this disease. Therefore, it is very important to find effective and safe neutralizing antibodies and vaccines for COVID-19. The RDB section of the spike protein is a suitable option for the production of subunit vaccines and neutralizing antibodies. This research aims to introduce the RBD section as a vaccine candidate.

The RBD was recombinantly expressed in two hosts, Escherichia coli (E.coli) and insect cells, and its production rate was compared. Using the western blotting method, protein production was confirmed. To evaluate the function of the protein expressed in two prokaryotic and eukaryotic hosts, using the serum of patients recovered from COVID-19 (wild type and delta), the ELISA method was used.

Despite the higher production of recombinant protein by E.coli, the affinity of the antibodies in the serum of the patients to the protein expressed in the insect cell was higher.

In this study, the RBD was expressed in two different hosts. The results show that RBD is mentioned as a vaccine candidate. Homozygous in the insect cell preserves the biological activity of the protein by carrying out the process of post-translational changes such as glycosylation.

High efficiency and purity protein purification is a serious challenge in the production of recombinant proteins. This study is proposing an easy and new strategy for fabricating and describing magnetic microspheres consisting of a silica-coated magnetic core and a polyvinyl acetate shell modified with iminodiastic-acetic acid and complexed with Ni2+, (Fe3O4@SiO2@PVA@IDA-Ni2+) for the separation and purification of organophosphorus hydrolase (OPH) protein and histidine-containing outer membrane protein (OmpW).

The nanoparticles were synthesized in the following four steps: 1) synthesis of nanoparticles with vinyl groups on their surface, 2) grafting of polyvinyl alcohol (PVA) to these particles, 3) conversion of the hydroxyl groups of PVA to iminodiacetic acid, and 4) charging with nickel ions to form chelate groups. Structural properties and physicochemical parameters of nanoparticles were determined by FTIR, DLS, SEM, TEM, EDX, TGA and VSM analyzes. Recombinant protein subunits OPH and OmpW were expressed and purified as recombinant and their structure was confirmed. In this study, the genes encoding the recombinant proteins OPH and OmpW were cloned into the expression vector pet-26b (+) and expressed in E. coli BL21 (DE3) as a host and secreted into bacterial culture medium. Gene expression and purification were assessed by sodium dodecylsulfate-polyacrylamide gel (SDS-Page and Bradford test).

The results confirm that uniform and spherical magnetic polymer nanoparticles with high magnetization and superparamagnetic properties were successfully synthesized. According to the SEM images, the average diameter of the nanostructure was observed in the range of 24.56-62.19 nm. The formation of Fe3O4@SiO2@PVA/IDA-Ni2+ nanoparticles was confirmed by the peaks identified at 1405 and 1600 Cm-1 corresponding to the symmetric and asymmetric stretching vibration of the O=C−O− group and the removal of the peak at 1740 Cm-1. These microspheres showed a high degree of recovery of protein bands regardless of the nature of the protein. OmpW and OPH proteins as a His-tagged recombinant protein were expressed and purified using the final nanoparticles.

According to the SDS-PAGE results, there was a high degree of resolution in protein separation. The synthesized nanoparticles have a high protein binding capacity of about 1.25 mg of protein per mg of nanoparticles.

Urinary tract infections have significant complications and high costs and can lead to death. Appropriate treatment has been challenged due to high drug resistance. The aim of the present study was the biosynthesis of zinc oxide nanoparticles/banana peel on multi-drug resistant uropathogenic bacteria in children under two years of age.

In this study, bionanoparticles of zinc oxide/banana peel were synthesized and the structural, optical, morphological, and particle size characteristics were investigated using X-ray diffraction, ultraviolet absorption spectrum, and scanning electron microscopy. Biochemical tests and Colony-PCR technique were used to identify bacteria. The antibacterial properties of the synthesized nanoparticles were evaluated by the Agar dilution method on isolated bacteria.

According to the XRD, SEM, and UV-Vis results, the synthesized zinc oxide had a hexagonal structure with a size of 100 nm and absorption at a wavelength of 258 nm. Bacterial strains of Escherichia coli, Proteus mirabilis and Klebsiella pneumoniae multidrug-resistant Uropathogenic bacteria were identified using morphological, biochemical, and molecular characteristics. The results of antibacterial tests showed that bionanoparticles of zinc oxide/banana peel at a concentration of 0.6 g/L were effective against all bacteria.

The findings of this research showed that zinc oxide/banana peel bio-nanoparticles had an effective antibacterial effect against multi-drug resistant UTI bacteria in children under two years of age.

Plants are a rich source of bioactive chemical compounds that play an important role in the prevention and treatment of infectious diseases. In this study, the ability of aqueous and ethanolic extracts of sumac (Rhus coriaria) fruit to inhibit the growth of Trichomonas vaginalis and Candida albicans were investigated in vitro.

Extraction of sumac fruit was done by soaking and stirring with water and ethanol and its antimicrobial effects were evaluated by Broth microdilution method by determining the minimum inhibitory concentration (MIC).

The 50% inhibitory concentration (IC50) of ethanolic extract against Trichomonas vaginalis was 1.8 mg/ml and for aqueous extract was 3.55 mg/ml in 24 hours. No concentration of the aqueous extract of Sumac fruit was able to inhibit the growth of Candida albicans, while the ethanolic extract at 60 mg/ml inhibited the growth of Candida albicans after 24 hours.

Sumac fruit extracts can be considered as a potential alternative to chemical drugs in the treatment of Trichomonas vaginalis infections, but these compounds have not shown favorable antifungal results against Candida albicans.