Journal of Cell and Molecular Research
Volume:14 Issue: 2, Winter and Spring 2022

In recent years, various efforts have been made to improve the functional potency of cancer drugs. Due to the fact that microRNA in the cell can act as a tumor inhibitor, it can be used as a suitable treatment for most cancers. In this study, chitosan coating is considered a factor that enhances function and effectiveness of microRNAs. MCF-7 cell line was divided into four experimental groups, including an untreated MCF-7 cell group, MCF-7 cell group with miR encapsulated with chitosan, MCF-7 cell group with chitosan, and MCF-7 cell group with doxorubicin as a positive control group. The effect of different concentrations of miR-372 was first evaluated, and the optimal dose was selected to evaluate the following parameters: the induction of cell death by applying flow cytometry, the cell survival by MTT, and the level of P53 protein by immunocytochemistry. The results showed that the dose of 1500 ng/μl of miR-372 coated with chitosan could induce cell death up to 50% in 24 and 72 hours of treatment. In addition, the rate of induction of cell death in the group treated with miR-372 coated with chitosan for 72 hours was statistically significant compared with the control group. In addition, the expression level of P53 protein in the same group was statistically significant compared with the control group. According to the results, the use of cell proliferation cycle regulators such as miR-372 can control the process of proliferation and thus improve the treatment of cancer.

Endophytic fungi are often producing host plant metabolites. Tanshinones are secondary metabolites of the Salvia genus which are also produced by some endophytic fungi. Efficient secondary metabolite production in endophytic fungi drops significantly after sequential subcultures. 5-azacytidine (5-AC) is an analog of the naturally occurring pyrimidine nucleoside cytidine and a DNA methyltransferase inhibitor. In this relation, 5-AC is an effective tool to induce the expression of silenced secondary metabolite genes in fungi. We isolated 4 endophytic fungi from the roots of Salvia abrotanoides which produced tanshinone. Cryptotanshinone and tanshinone IIA were produced by Penicillium canescens, Penicillium nodositatum, and Penicillium pinophilum, while Paraphoma radicina only produced tanshinone IIA. The maximum amount of tanshinones was extracted from P. pinophilum culture with 130.826 mg cryptotanshinone /g of dry weight and 50.155 mg Tanshinone IIA/g of dry weight. These amounts were significantly more than tanshinones produced in plant roots (0.55 mg cryptotanshinone/g of dry weight, 1.3 mg Tanshinone IIA/g of dry weight). In the third subculture, tanshinone production decreased significantly. 5-azacytidine as an epigenetic modifier retrieved tanshinone production in the third subculture of P. pinophilum. Also, 5- azacytidine treatment made a big jump in Tanshinone IIA production in P. radicina (63.176 mg TIIA/g of dry weight) besides increasing Tanshinone IIA production in P. nodositatum cultures. This is the first report using 5- azacytidine to improve tanshinone production in endophytic fungi. Our results confirm that 5- azacytidine is an efficient, easy, and quick chemical to felicitate secondary metabolite production in endophytic fungi.

Today, ostrich breeding has been widely developed in Iran and other countries due to the ability of this animal to produce quality meat, leather, and oil. However, one of the main problems in breeding them is sex determination using aggressive techniques with low accuracy. This study aimed to determine the sex of immature ostriches using specific primers in a multiplex PCR reaction. This study considered  20 specimens of unspecified immature and six specimens (three adult males and females) of known-sex African ostriches as controls. SS and OSFES primers were used to amplify part of the female-specific sequence and 18S primer was used as a control in a PCR reaction. The presence of SS and OSFES bands in gel electrophoresis indicated the amplification of the desired parts related to the female sex and the absence of these bands indicates the male sex of the species. In total, out of 20 African ostriches studied, 50% of them belonged to females and 50% of them belonged to males. Later, with the growth of immature individuals, the results of this experiment were confirmed. In this study, it was found that the use of feather samples for DNA extraction and multiplex PCR is a suitable, accurate, and cost-effective method in identifying and determining the sex of young ostrich and leads to more real and reliable results, avoiding stress in birds.

Spinal cord injury (SCI) is a severe central nervous system trauma (CNS) that has two primary and secondary phases. The initial phase, which is irreversible, causes nerve tissue destruction and bleeding. Various factors in the second phase together aggravate the primary damage. One of the important factors of the second phase is the cascading of inflammatory factors, which, contribute to the further destruction of nerve tissue. In addition to surgical treatments, drug and cell-based or extracellular vesicles therapy, by modulating the immune system and reducing inflammatory factors at the lesion site, prevent further destruction of nerve tissue and help improve the patient's neurological and motor function. Researchers have provided many chemical and herbal medicines to reduce complications caused by spinal cord injury, many of which are currently being used and are also known as drugs of choice. However, sometimes the long-term use of these drugs causes side effects. Today, the new approach of cell therapy and the use of extracellular vesicles (EVs) is being investigated, which has minimized the side effects of drug treatments and helped to improve the function of nerve cells. Mesenchymal stem cells (MSCs), have a high ability to differentiate into different cells and to modulate the immune system by secreting paracrine factors. But since they cannot cross the blood-brain barrier (BBB), researchers solved this problem by extracting extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs), which also contain all paracrine factors. In this study, a brief overview of drug treatments, stem cells, and extracellular vesicular therapy in the treatment of spinal cord injury has been discussed.

Gastric carcinoma (GC) is the third leading cause of malignancy-related deaths worldwide, and Helicobacter pylori is one of the identified causes of gastric cancer. The Cag pathogenicity island (cagA, cagC, virB2) is one of the major pathogens of H. pylori, which increased the risk of gastric cancer development. Some studies have shown that H. pylori significantly alters the expression of certain genes in gastric epithelial cells. This study aimed to investigate the expression of the GPR83, CA1, AWP1, and WTAP genes in AGS cells transfected with the recombinant pIRES2-EGFP-cagC vector. The pIRES2-EGFP-cagC and pIRES2-EGFP plasmids (as controls) were transfected into AGS cells by lipofectamine 2000 solution. Then, RNA extraction and cDNA synthesis were performed. The expression of GPR83, CA1, AWP1, and WTAP genes was evaluated using the real-time PCR method. Finally, the expression of each gene was evaluated using SPSS software and t-test independent statistical tests. Our findings indicated that the expression of the GPR83 gene in AGS cells treated with cagC statistically significantly increased compared to control cells (P=0.0327). On the contrary, the WTAP expression was significantly decreased (P=0.0132), whereas the AWP1 and CA1 mRNA expression levels were not statistically significant. This study shows that GPR83 and WTAP genes's expression in the host is significantly altered through cagC gene expression. Hence, it seems that the cagC gene's presence can explain some alterations in the expression of gastric epithelial cell genes and the cause of gastric cancer pathogenesis caused by H. pylori infection.

Prostate cancer is the second most prevalent malignancy in men and the fifth leading cause of death worldwide. It is the second most common urinary tract cancer among Iranian men. The aim of this study was to investigate the association between rs10090154, located on the 8q24 locus, and rs1691053 on chromosome 5p15.31, with prostate adenocarcinoma and PSA levels. This study also aimed to identify the potential of these genetic markers as screening factors. This case-control study included 79 patients with prostate adenocarcinoma aged between 48 to 86 years, as well as 98 patients with benign prostatic hyperplasia aged between 47 to 81 years. The Tetra-primer ARMS-PCR method was applied to determine the genotype of each participant. In this study, no significant differences in genotypic distribution between the prostate adenocarcinoma and control groups were discovered for rs10090154 (P-value = 0.608) and rs1691053 (P-value = 0.102) polymorphisms. Moreover, for the study of the additive genetics model of the rs10090154 polymorphism, with the TT genotype as the reference, the CC genotype with P-value = 1 and OR {95%CI} = 0.750, {0.039-14.576} and CT genotype with P-value = 0.324 and OR {95%CI} = 0.577, {0.191-1.739}, were not associated. Correspondingly, for rs1691053, with the CC genotype as the reference, the CT genotype with P-value = 0.176 and OR {95%CI}= 0.196, {0.022-1.793}, and the TT genotype with P-value= 0.464, OR{95%CI}= 0.125, {0.005-3.225}, were not associated. These findings suggest that rs10090154 and rs1691053 may not be associated with prostate cancer among Iranians. However, further research with larger sample sizes and investigation of various Iranian subpopulations are needed to confirm these results.

Prostate cancer is one of the leading causes of cancer-related deaths among men worldwide. Research has shown that genetic variations can increase an individual's risk of developing this disease. In this study, we investigated the potential role of two genetic variants, one within RFX6 and another within SLC22A3, in predisposition to prostate cancer. A genetic association study was conducted, involving a case-control design with 112 prostate cancer cases and 95 individuals affected by benign prostatic hyperplasia who served as controls and had no history of cancer. The genotypes of the two variants, rs339331 in RFX6 and rs9364554 in SLC22A3, were determined using tetra-primer ARMS-PCR. In this study, a multi-stage strategy was employed to analyze the data obtained from genotyping and to assess the association of these two variants with prostate cancer risk. For statistical analysis, Chi-squared, Fisher’s exact test and logistic regression were performed to evaluate the association of variants with prostate cancer and Gleason score. The results suggest that the rs9364554 variant in SLC22A3 is not associated with prostate cancer risk under either additive or multiplicative genetic models, while the variant in RFX6 is significantly associated with prostate cancer susceptibility. The TT genotype of rs339331 indicated a possible protective effect against prostate adenocarcinoma (CI 95% = 0/009 -0/725, P-value = 0/009, OR = 0/083). In conclusion, our study provides evidence that the genetic variant rs339331 within RFX6 is significantly associated with prostate cancer susceptibility and may have a potential protective effect against prostate adenocarcinoma. It should be mentioned that more research is needed to investigate the possible protective effect of the TT genotype of rs9364554 against prostate cancer risk in other populations and various ethnic groups and particularly larger sample sizes are required to confirm this association.

Cytochromes are enzymes of the dehydrogenase class with a hemoprotein structure in which the iron in these compounds undergoes oxidation and reduction reactions upon receiving or losing electrons. Quantitatively speaking, CYP3A4 is the most important isoenzyme of cytochrome P450, oxidizing foreign organic molecules such as drugs or toxins to cause them to leave the body. Many drugs and antibiotics can induce or inhibit the activity of cytochrome P450, including dexamethasone. Dexamethasone is a steroidal anti-inflammatory drug used to treat of inflammatory diseases and chronic autoimmunity. This study aimed to investigate the induction effect of dexamethasone in biotransformation pathways by in silico tools. Molegro Virtual Docker software was used to investigate the molecular docking of the enzyme and dexamethasone, which indicated the binding of the drug to the enzyme.  The molecular simulation was performed in Linux with the GROMACS program.  Root-mean-square distance (RMSD), and radius of gyration (Rg), were evaluated. The results were analyzed with Pymol and VMD software, and the obtained curve was plotted with GRACE software. Docking results show that a cluster with a bond energy of -60.81 was the best cluster, and the bond size between ligand and internal atoms was -23.191in the complex. In addition, the amount of bond between the ligand and water for this pose was zero. The stability of the enzyme-ligand complex and the induction effect of dexamethasone on CYP3A4 were indicated by RMSD and RG results. Results of RMSD and RG of CYP3A4 glucocorticoid obtained from the simulation showed the stability of binding of the drug to the enzyme. Also, RMSD results showed the stability of glucocorticoid and dexamethasone complex during molecular dynamics simulation. It reached relative stability at 0.8 nm after 80,000 ps until the end of the simulation.