Cell and Tissue Journal

Abstract Introduction: Concerns about fossil energy costs, environmental deterioration, and energy security has created strong motivation for the research and development of routes to provide sustainable and renewable fuels. In recent years, the use of biomass to produce highly valued chemicals has attracted widespread attention. Lignocellulosic biomass, as a promising renewable resource for biofuel production, has distinct advantages in terms of economic and environmental benefits. The conversion of renewable raw materials to hydrocarbon fuels is an attractive alternative to fossil fuels from economic and environmental perspectives. The production process of lignocellulosic biomass mainly consists of biomass accumulation, biomass decomposition, simple sugars, and conversion of sugars to biofuel. One of the crucial steps for the economic success of lignocellulosic biofuels depends on the inhibition of competitive metabolism in microorganisms to achieve high productivity. To date, there has been a growing focus on the use of S. cerevisiae and E. coli as cell lines. These two cellular factories have well known advantages. They are genetically transmissible and several tools are available for genetic manipulation. In order to produce xylonate, the engineered xylose is first converted by a dehydrogenase into the intermediate xylonolactone, which is then slowly converted to xylonate in a nonenzymatic reaction.
Aim: The organic compound D-1,2,4-Butanetriol (BT) is a valuable chemical with wide-ranging applications in various fields such as pharmaceuticals, paper, polymer materials, and military applications. However, the chemical synthesis routes for BT have many drawbacks. By genetically modifying microorganisms, the metabolic pathway for producing many substances, including BT, can be engineered. When D-xylose is supplied to the bacterium, it is first converted into an intermediate compound called xylonolactone. This compound slowly converts into xylonate through a non-enzymatic reaction. To produce xylonate, the engineered bacteria receive xylose, which is initially converted by a dehydrogenase reaction catalysed by the xylose dehydrogenase enzyme into an intermediate compound, xylonolactone. Xylonolactone is slowly converted to xylonate in a nonenzymatic reaction. Xylonate is a five-carbon organic acid. Over the past few years, xylonate has increasingly been considered as an important chemical due to its potential as an important chemical component. Xylonate has many applications in the food, chemical, and pharmaceutical industries. Specifically, xylonate can act as a precursor for the synthesis of D-1,2,4-Butanetriol and as a concrete water reducing agent. E. coli was chosen as the target strain for genetic and metabolic engineering due to its fast growth in inexpensive culture media, the presence of two enzymes for BT synthesis, and product formation in less than 24 hours of fermentation. This study aimed to clone and express xylose dehydrogenase from Caulobacter vibrioides in E.coli.
Materials and Methods: At first, to access the bacterial gene sequence, the genome of the target bacterium was extracted. Then, to create a strain expressing the enzymes xylose dehydrogenase and xylonolactonase, the genes for these proteins were amplified from Caulobacter vibrioides CB1 and transferred into E. coli. For this purpose, the target genes were amplified using specifically designed primers via the Polymerase Chain Reaction (PCR) method and initially cloned into a pTZ57cloning vector and then subcloned into pET 26b expression vector. At the final step, the expression of the enzyme was assessed by SDS-PAGE, and the other confirmation was the reduction of NAD+ to NADH, which was used as an activity indicator of the enzyme, as investigated by a change in NADH absorbance at 340 nm.
Results: Confirmatory tests were performed to ensure the presence of the gene in the vectors (using restriction enzymes and colony PCR for gene amplification). The expression and activity of the enzyme were analyzed. The recombinant protein's presence was confirmed by SDS-PAGE for the xylose dehydrogenase gene, with a molecular weight of 52.2 kDa. The estimated expression level of the recombinant protein was approximately 25%.
Conclusion: The objective of this research was solely to establish the metabolic pathway for xylonate production in E. coli by surface expression of enzymes in this pathway (xylose dehydrogenase). The results obtained in this study confirm that half of the pathway is active at the cell surface, but further experiments are required to determine the precise production levels and complete the pathway.

Abstract Introduction: Pancreatic cancer is the fourth leading cause of cancer-related deaths worldwide. 5-Fluorouracil is one of the commonly used chemotherapeutic drugs. The plant Artemisia absinthium has attracted attention as a potential herbal anticancer agent. This study investigated the effect of the methanolic extract of this plant on the expression of miR-34a and the apoptotic genes BAX, Caspase-3, and Caspase-9 in AsPC-1 pancreatic cancer cells.
Aims: This study aimed to evaluate the cytotoxic and pro-apoptotic effects of Artemisia absinthium methanol extract on pancreatic cancer cells. The research specifically investigated the molecular mechanism by analyzing expression changes in the tumor suppressor miR-34a and key apoptotic genes BAX, Caspase-3, and Caspase-9 to elucidate the extract's anti-cancer mode of action.
Materials and methods: To evaluate the cytotoxic effects of the plant extract on cancer cells, an MTT assay was performed to determine the viability and survival rate of the cells following treatment with various concentrations of the extract, the chemotherapeutic drug fluorouracil (5-FU), and the combined treatment of the extract and the drug. This assay measures cellular metabolic activity and allows quantification of live and dead cells after exposure to different treatments. Based on the obtained results, the IC₅₀ value for each treatment was calculated, representing the concentration at which 50% of the cells were inhibited or killed.
After determining the IC₅₀ value, cells were treated with concentrations equal to, lower, and higher than the IC₅₀ to further investigate the cytotoxic effects and the mode of cell death induced by the treatments. To distinguish between apoptotic and necrotic cell death, the Annexin V-FITC/PI assay was employed. This assay detects phosphatidylserine externalization on the cell membrane and enables differentiation between live, early apoptotic, late apoptotic, and necrotic cells.

In addition to the morphological and physiological assessments, molecular analyses were conducted to examine the expression levels of key apoptosis-related genes, including Caspase-3, Caspase-9, and BAX, as well as the regulatory microRNA miR-34a. Gene expression analysis was performed using Real-time PCR (qPCR).

Results: The results of the MTT assay demonstrated that the proliferation of AsPC-1 pancreatic cancer cells was inhibited by treatment with the extract of Artemisia absinthium and the chemotherapeutic drug fluorouracil (5-FU) in a concentration-dependent manner. As the concentration of each treatment increased, cell viability significantly decreased, indicating a marked cytotoxic effect of both the plant extract and the drug. Moreover, a possible synergistic effect between the extract and fluorouracil in suppressing. To determine the mode of cell death induced by these treatments, the Annexin V-FITC/PI assay was performed. The results revealed that a considerable proportion of treated cells underwent programmed cell death (apoptosis), while the percentage of necrotic cells remained relatively low. These findings suggest that the observed reduction in cell viability is mainly mediated through the activation of apoptotic pathways rather than necrosis. Furthermore, Real-time PCR analysis showed a significant upregulation in the expression of the regulatory microRNA miR-34a and the apoptosis-related genes Caspase-3, Caspase-9, and BAX in the treated groups compared to the control group.
Discussion: The obtained results suggest that Artemisia absinthium extract exerts its cytotoxic effect primarily through the induction of apoptosis rather than necrosis in AsPC-1 pancreatic cancer cells. The observed upregulation of miR-34a, Caspase-3, Caspase-9, and BAX implies activation of intrinsic apoptotic pathways. These findings are consistent with previous studies reporting pro-apoptotic properties of A. absinthium and other Artemisia species. Therefore, the extract may enhance the therapeutic response of pancreatic cancer cells when combined with conventional chemotherapeutic agents such as fluorouracil.
Conclusion: In conclusion, Artemisia absinthium extract demonstrated strong antiproliferative and apoptosis-inducing effects on AsPC-1 cancer cells in a dose-dependent manner. Its combination with fluorouracil produced a synergistic cytotoxic impact, significantly enhancing cell death through apoptotic signaling. The molecular findings support the potential of this extract as a complementary therapeutic agent. Further studies are recommended to explore its mechanisms and evaluate its efficacy in in vivo models of pancreatic cancer.

Abstract Introduction: Breast cancer is the most important and widespread type of cancer in women's population.
Aim:This study was conducted to synthesize zinc oxide nanoparticles conjugated with gingerol (ZnO@CPTMS-Gingerol) and evaluate their anticancer effects on breast cancer cells.
Materials and Methods: To synthesize ZnO@CPTMS-Gingerol nanoparticles, one gram of ZnO@CPTMS nanoparticles was dispersed in 30 ml of dry toluene. One gram of gingerol and 10 ml of triethylamine were added to the reaction mixture and refluxed for 24 hours. The product was washed twice with a mixture of distilled water and ethanol (1:1), and the final product was dried at 100°C for 24 hours. Physicochemical properties of ZnO@CPTMS-Gingerol nanoparticles were studied by FT-IR, XRD, DLS, EDS, zeta potential measurement, and electron microscope imaging. The inhibitory effects of different concentrations of ZnO@CPTMS-Gingerol nanoparticles on MCF-7 breast cancer cells and HEK293, as normal cells, were evaluated by the MTT test. To perform this experiment, cells were prepared in 96-well cell culture plates with a density of 104 cells/well, and then were treated with concentrations of 15.625, 31.25, 62.5, 125, 250, and 500 μg/mL of ZnO@CPTMS-Gingerol. After incubating the cells for 24 hours at 37°C, 0.2 ml of MTT solution was added to each well. The wells without nanoparticles treatment were considered as controls. After incubation for 4 hours, the supernatant was removed, and 100 μl of DMSO solution was added to each well. After pipetting, the optical density was read at 570 nm using an ELISA Reader. To determine the percentage of apoptotic and necrotic cells, 5x10⁵ cells were treated with ZnO@CPTMS-Gingerol nanoparticles for 24 hours with half inhibitory concentration (IC50). Then, the treated and control cells were stained with annexin V and propidium iodide (PI) dyes. Finally, cell analysis was done by a flow cytometer. Data analysis was done using device software and dividing the points recorded in the two-dimensional curve into four regions including Q1 to Q4. The experiments were performed in three replicates, and the results were expressed as mean ± standard deviation. Statistical analysis including t-tests, and one-way ANOVA was performed using SPSS. A p

Abstract Aims: Nanoparticles due to their wide applications in medicine,industry,and biotechnology, have attracted many scientists’ attentions. Recently, nanoparticles especially selenium nanoparticles are widely used to diagnosis and cancer treatment. The aim of this study was to evaluate the cytotoxic and anticancer effects of selenium nanoparticles on colon cancer cell line and analysis of CAD (Caspase Activated DNase) gene expression.
Material and methods: In this study, colon cancer HT29 and normal HEK293 cell lines were purchased from the Pasteur Institute Cell Bank of Tehran and treated with selenium nanoparticles overnight. The cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Scotland) medium with 10% FBS serum and 1% streptomycin antibiotic (Gibco, Scotland). The cells were then stored at 37 ° C. In this study, cytotoxic effect of Selenium NPs was evaluated on HT29 and HEK293 cells using MTT (3-(4, 5-Dimethyltetrazollium Bromide) assay. Subsequently, they were treated with selenium nanoparticles in different concentrations (0, 7.81, 15.62, 31.25, 62.5, 125, 250 and 500 mg/mL) for 24 hours. To solubilize the viable cells formazan crystals production, we added 100 μl/well of dimethyl sulfoxide (DMSO) to them. After treatment of HT29 cells with IC₅₀ concentration, the total RNA was extracted and cDNA synthesized. Moreover, CAD gene expression was evaluated using Real Time PCR method. The data was evaluated by ABI StepOne utilizing the Applied Biosystems qRT-PCR (ABI 7300 system, Applied Biosystems). The quantification of the mode of Selenium NPs -induced cell death in the HT29 cells were ascertained using flow cytometry followed by staining with fluorescein isothiocyanate (FITC)‐Annexin V and propidium iodide (PI) staining. Finally, the study of apoptosis and necrosis of Selenium NPs was evaluated using flow cytometry method. Data analysis was statistically determined by using One-way analysis of variance (ANOVA) with SPSS/22 software followed by a Tukey test.
Results: The result showed that the treatment of Selenium NPs at 31.25 to 500 µg/mL concentration had maximum cytotoxic effect, revealed statistically significant (P˂0.001). The IC₅₀ value for Selenium NPs were measured at 75 µg/mL after 24 hours. In order to determine the effect of Selenium NPs on cancerous cells, alterations in the mRNA expression levels of CAD gene in HT29 cells were done by qRT-PCR technique followed by the exposure to nanoparticle. The CAD gene expression comparing to reference gene was up-regulated 4.04±0.125 fold. To determine the mechanism of cell death in the cancer cells, annexin V/PI flow cytometry was carried out. In the treatment of HT29 cells by IC₅₀ of selenium NPs, 10.43%, and, 24.28% of early and late stages’ apoptosis were observed, respectively
Conclusion: Our results suggest that selenium NPs can display some promising cytotoxic properties through inducing apoptosis pathway. Based on the results, up-regulated gene expression involved in apoptosis (CAD) and activating apoptosis, it can be concluded that the selenium NPs can be used as drug candidate in colon cancer treatment, but more studies are needed regarding the medicinal importance of nanoparticles.

Abstract Aim: Organotypic cultures of spinal cord slices from mammalian neonatal and fetal animals are powerful tools for studies of spinal cord injury, neuronal degeneration, and cell death but also motor neuron regeneration. Models in which adult slices are used would be very useful. However, adult spinal cord slices are notoriously difficult to maintain in culture and rapidly deteriorate in vitro. Degeneration of motor neurons in the spinal cord is a critical phenomenon in spinal cord injuries and certain neurodegenerative diseases such as amyotrophic lateral sclerosis, a neurodegenerative disorder in which motor neurons in the spinal cord and motor cortex are lost. A variety of mechanisms have been proposed as having a role in neuronal apoptosis during spinal cord injury. Oxidative stress has been reported as one of the mechanisms involved in the apoptosis of motor neurons in spinal cord injuries and neurodegenerative diseases. This study was conducted to determine whether quercetin, as a potent antioxidant, can delay apoptosis in motor neurons of cultured spinal cords by reducing oxidative stress.
 Material and methods: The thoracic regions of the spinal cord from adult NMRI mice were sliced using a tissue chopper and divided into three groups: 1) 0-hour, 2) control group, and 3) group treated with quercetin (100 µM). Spinal cord slices in the 2 and 3 groups were incubated for 6 hours at 37°C in a Co₂ incubator. The viability of the spinal cord slices was measured using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The morphological features of apoptosis and the number of motor neurons were examined using Hoechst and propidium iodide staining. The malondialdehyde was measured to determine lipid peroxidation while the FRAP (ferric reducing antioxidant power) was assessed to evaluate total antioxidant capacity in fresh and cultured spinal cord slices. Results were expressed as mean±SD. One-way analysis of variance (ANOVA) followed by Tucky’s test was used to assess the statistical significances of the data. In all cases, a statistical probability of p<0.05 was considered significant.
Results: After 6 hours (control group) in culture, the viability of the spinal cord slices, the neuron diameter, and the number of healthy motor neurons significantly decreased compared to the 0-hour group. Also, motor neurons showed the morphological features of apoptosis including cell shrinkage, nuclear and chromatin condensation in the control group. A significant increase in the amount of malondialdehyde and a significant decrease in the total antioxidant capacity was also observed compared with the 0-hour group. After 6 hours, quercetin not only increased the viability and the number of healthy motor neurons in the cultured slices but also reduced the morphological features of apoptosis in the motor neurons compared with the control group. In addition, quercetin significantly reduced the amount of malondialdehyde and increased the total antioxidant power in slices cultured for 6 hours.
Conclusion: Oxidative stress might be considered as one of the mechanisms involved in the apoptosis of motor neurons in cultured spinal cord slices and quercetin, as a potent antioxidant, was able to increase the viability of the cultured spinal cord slices and delay the morphological features of apoptosis in the motor neurons through reducing lipid peroxidation and increasing the total antioxidant capacity.

Abstract Aim: Cancer remains a global health problem, with ovarian cancer ranking fifth among cancers affecting women and the leading cause of cancer-related death in women. There are different types pf ovarian cancer, including Epithelial ovarian cancer, Stromal tumors and Germ cell tumors.  Epithelial ovarian cancer is the most common type that includes several subtypes, including serous carcinoma and mucinous carcinoma. To this end, several factors have been identified to increase your risk of ovarian cancer, including older age, inherited gene changes, family history of ovarian cancer, being overweight or obese, postmenopausal hormone replacement therapy, endometriosis, and never having been pregnant. The current strategies to treat ovarian cancer include surgery, chemotherapy, radiotherapy and targeted therapies, and hormone therapy. Scientists continue to investigate the foundational mechanisms involving cancer development, as well as to find new drugs and metabolites having the capacity to control cancer. Alpha-ketoglutarate (AKG), a critical metabolite in the Krebs cycle involved in cellular energy production and the regulation of gene expression. Recent studies have shown that AKG may have the potential to enhance the efficacy of cancer treatments, by modulating the tumor microenvironment and improving the immune response against cancer cells. This study investigates the effect of AKG on ovarian cancer cells.
Material and methods: SKOV3 cells were obtained from Tehran University and cultured in complete culture medium (RPMI, 10% Fetal bovine serum (FBS), and 1% Penicillin-Streptomycin (Pen/Strep)). To find the proper concentration of AKG, SKOV3 cells were cultures in 96-well plate, and treated with different concentration of AKG (range 20 to 220 µM). After 24 and 48 h, the viability of the cells was determined using MTT assay. Based on the results obtained from viability assay, 200 μM of AKG was selected for the next assessments. To evaluate the effect of AKG on SKOV3 cell proliferation using plotting a growth curve, cells were cultured in the presence (200 μM AKG) and absence of AKG, and counted the number of cells every 24h for one week. To determine the population doubling time (PDT), the cells were cultured in the presence (200 μM AKG) and absence of AKG for 72h, then the cell were collected and the number of living cells was counted using Neubauer Chamber. The PDT was calculated using a related standard method. To assess colony formation potential, the SKOV3 cell were cultured in the presence (200 μM AKG) and absence of AKG. After 7 days, the cells were fixed using 10% formalin solution, then the colonies were stained using crystal violet dye, and the number of colonies were counted using inverted microscope. To investigate the effect of AKG on the migration rate of SKOV3 cells, the cells were cultured in complete medium to reach 85% confluence, then treated with mitomycin (10 μM) for 3h, then Created a scratch in the cell monolayer using a sterile pipette tip. the cells were cultured in the presence (200 μM AKG) and absence of AKG for 3 days. The images of the scratch were taken at regular intervals using a microscope, and the closure of the scratch over time was analyzed. To analyze the cell cycle profile, the SKOV3 cell were cultured in the presence (200 μM AKG) and absence of AKG for 48h. Then, the cells were collected and analyzed using a flow cytometry.
Results: Based on the MTT assay, 200 μM AKG was determined as the proper concentration to investigate other biological behaviors of SKOV3 cells. Colony forming assay showed a decrease in the number and size of colonies in the AKG-treated group (P<0.05). In addition, the cell doubling time increased in the treatment group, indicating slower growth rate (P<0.05). Growth curve analysis confirmed reduced cell growth in treated group. Cell cycle analysis showed a higher percentage of treated cells arrested in S and G1 phases. The scratch assay showed slow cell migration and metastasis in the cells treated with 200 µM AKG.
Conclusion: In conclusion, AKG has an inhibitory effect on the proliferation, viability, migration in SKOV3 ovarian cancer cells, highlighting its potential as an adjuvant treatment with existing therapies. More research is necessary to fully investigate the therapeutic effect of AKG in ovarian cancer.
 

Abstract  
Aim: Optimizing cell culture conditions to improve oocyte growth and maturation in vitro culture conditions has been the focus of many researchers today, but the quality improvement mechanism is not fully understood. Apoptosis or programmed cell death is a process to remove old and damaged cells from tissues and plays a major role in the life of follicles and immature oocytes. Most of the defective reproductive cells as well as extra cells are removed from the ovaries through apoptosis. One of the ways to reduce oxidative stress in the laboratory culture of oocyte maturation is the use of antioxidants. Statins are drugs for the treatment of high cholesterol for which antioxidant and anti-apoptotic properties have been reported. Atorvastatin in high doses has side effects for the heart and kidneys, but in low concentrations, it has antioxidant and anti-inflammatory effects for cells, and no side effects have been reported for it.In this study, we investigated the effect of low-dose atorvastatin on the rate of apoptosis in mouse oocytes.
 
Materials and Method: 24 h after PMSG (Pregnant Mare Serum Gonadotropin) inhection, 200 oocytes were obtained from adult female Wistar rats at the age of 4-5 weeks, were from were divided into 2 groups of 100 including the control group (MEM:Minimum Essential Medium culture+ Growth factor) and the atorvastatin group (MEM culture medium + 2 mg/kg atorvastatin+Growth factor) and cultured in the incubator(35 ^oC,CO2 %). After 24 hours of oocyte culture, Matured oocytes that met the standards of a healthy mature oocyte(oocytes that were in the first meiosis and had the first and mature polar body) were isolated the amount of apoptosis in each group was checked using tunnel staining and fluorescent microscope, the data were analyzed by variance analysis.
Results: After 24 hours of oocyte culture, the amount of apoptosis in the group receiving atorvastatin in the culture medium and the control group was investigated. The rate of apoptosis in the atorvastatin group and the control group was 24% and 22%, respectively, In the atorvastatin group, apoptosis increased by 2% compared to the control group butt the difference between the two groups was not statistically significant (p =0.11).
Conclusion: According to the findings of this study, atorvastatin in low doses can have a pro-apoptotic effect and cause partial induction of apoptosis in mouse oocytes in a laboratory environment. Although this study was not conducted on other doses of atorvastatin, it is suggested that the effect of other doses of this drug on the rate of apoptosis induction should be investigated in order to make an evidence-based decision regarding its administration and use.
 

Abstract Aim: The toxic effects of cadmium exposure are mainly due to the production of oxygen free radicals, reduction of cellular antioxidants and oxidative stress, which can lead to cell component destruction, DNA damage, apoptosis and ultimately tissue damage. In this study, the protective effects of N-acetylcysteine, as a substance with antioxidant properties, in cadmium-exposed Wistar mice were investigated by liver histology and measuring the expression of effective genes in apoptosis and cell proliferation.
Material and Methods: Mice weighing approximately 150-200 g were classified into three treatments including, G1) control treatment, G2) cadmium recipient treatment, and G3) concomitant cadmium and N-acetylcysteine treatment and for four weeks received the desired. Then liver tissue samples were taken for histopathological examination and expression of eif4e and mad1 genes.
Results: The results of this study showed that cadmium exposure resulted in serious damage to rat liver tissue, including central artery hyperemia, increased number of inflammatory cells and inflammation in the liver parenchyma, while the use of N-acetylcysteine significantly reduced the mentioned injuries. Also, the use of N-acetylcysteine resulted in a significant reduction in the expression of eif4e and mad1 genes by 2.14 and 2.27 times, compared with mice that received only cadmium.
Conclusion: The results of this study suggest that N-acetylcysteine as an antioxidant can play an important role in preventing tissue damage due to oxidative stress and preventing the induction of cellular apoptosis.

Abstract Aim: One of main challenges in worldwide is increasing rate of depression and sexual dysfunction associated with antidepressant drug consumption. Regarding the role of Sertoli cells in spermatogenesis, this study investigated the effect of duloxetine on viability, apoptosis and expression of Bax and Cx43 (Connexin 43) expression in Sertoli cells.
Material and Methods: TM4 Sertoli cells were cultured in DMEM/F12 medium containing 2.5% FBS, 5% horse serum and 1% penicillin-streptomycin. Cells were treated with different doses of duloxetine (3.75, 7.5, 15, 30, 60 µg/ml) for 24 to 72 hours. MTT assay was performed to evaluate cell viability. The rate of apoptosis was measured by flow cytometry and RT-qPCR was performed to evaluate of Bax (proapoptotic gene) and Cx43 (essential for spermatogenesis) genes.
Results: Duloxetine reduced cell survival in a dose and time-dependent manner. On the basis of

Abstract Aim: Cancer is one of the leading causes of death worldwide, and its treatment is always associated with side effects such as drug resistance. So, there is a strong tendency for the identification of new herbal anti-cancer compounds.
Material and Methods: In this study, a range of 0.5-12 µg/mL of hydroalcoholic extract of Tamarindus indica kernel and 5-Fluorouracil was applied to prostate cancer (LNCaP), colon cancer (HT-29) and normal fibroblast (HSkMC) cells for 24 hours. The cytotoxic effect of the extracts was measured by the MTT method. The rate of DNA synthesis and incidence of apoptosis was measured by BrdU and TUNEL assays, respectively.
Results: Following treatment with the highest amount of the extract, the viability of prostate, colon, and fibroblast cells was reduced to 4.8, 65.1, and 60.5%, respectively. The IC₅₀ for the three cell lines was 4.60, 17.0 and 13.79 μg/mL respectively. The rate of DNA synthesis also reduced by 32, 37 and 15% for prostate, colon and fibroblast cancer cells, respectively. The rate of apoptosis in LNCaP and HT-29 cells was 31 and 4%, respectively.
Conclusion: Collectively, the toxicity of the extract was higher for LNCaP cells than for the other two cells (p-value ˂0.01). Further studies in vivo and analysis of compounds in tamarind can lead to the identification of anti-cancer compounds from this plant.

Abstract

Abstract

Abstract

Abstract

Abstract Aim: The purpose of the current study is to investigate the cytotoxic and oxidative effects of Salvia officinalis hydroalcoholic extract on cancer A549 cells.
Material and Methods: The cells were seeded in plates to investigate concentrations of Salvia officinalis extract and MTT measurement was done to determine IC50 concentration and cell viability on days 1, 3, 5, and 7. Moreover, analyses of superoxide dismutase and catalase enzymes involved in oxidative stress pathway, and AO/EB staining for a qualitative investigation of cell lines has been conducted in order to explore the effects of IC50 concentration on apoptosis induction. Also, Giemsa and DAPI stainings were utilized to explore the morphological changes of cell and nucleus..
Results: IC50 concentration of Salvia extract for A549 cells was determined 5 mg/mL. According to the results, Salvia extract reduced the cell viability of A549 cells. Based on the results, Salvia extract had a significantly greater cytotoxic effect compared to the control sample of A549 cell line. Treatment cells indicated some clear and dose-dependent differences at different times. The results of stainings proved the apoptosis induction in the treatment group. Furthermore, regarding the enzyme expression results the activity of superoxide dismutase and catalase enzymes in the cell groups treated by Salvia extract was significantly increased, compared to the control group.
Conclusion: Consistent with the results of this study, in addition to the anti-oxidative activity Salvia officinalis hydroalcoholic extract revealed significant apoptotic effects on lung cancer cells, considering a time and dose-dependent method.
 

Abstract Aim: The aim of this study was to compare the anti-proliferative and apoptotic effects of hydroalcoholic extracts of different herbal medicines (Achillea wilhelmsii, Silybum marianumseed, Echinacea purpurea, Adiantum capillus-venerisand apricot kernel) against breast cancer cells (Mcf-7).
Material and method: For this purpose, the plants were dried and milled, then, soaked in 70% ethanol for 72 hours and their extracts were extracted using a rotary evaporator.Different concentrations of herbal extracts (12.5, 25, 50 and 100 μg/ml) were added to the cancer cell culture medium and their cytotoxicity and apoptotic effects were investigated after 24h by MTT assay and acridine orange - ethidium bromide staining, respectively.Data was analyzed by SPSS software at the significant level of 5%.
Results: Addition of the highestconcentration of all extracts to the culture mediumshowed the most significant anti-proliferative and apoptotic effects (p < 0.05) compared to other concentrations of the same extracts.Also, among the high concentrations (100μg/ml), the highest cell cytotoxicity effects were related to the extracts of Echinacea purpureaand Adiantum capillus-veneris (p < 0.05).
Conclusion: The results of this study indicated that the addition of Echinacea purpurea and Adiantum capillus-venerisextractsto the cell culturesin high concentrationshad the most significant anti proliferative and apoptotic effects on breast cancer cells in comparison with other plant extractsand concentrations.
 

Abstract Aims: This study was aimed to investigate the anti-proliferative effects of hydroalcoholic extract of Blepharis persica seed and its synergy effect with doxorubicin on human breast cancer and prostate cancer cell lines.
Materials and Methods: hydroalcoholic extract of  seed was prepared using maceration, and ethanol­%70. Eight concentrations of extract and four concentrations of combined with doxorubicin (125, 62.5ngr/ml) and extract (0.625, 0.315­mg/ml) were prepared. MCF7, LNCaP and SKM cell lines were cultured .Cell­­ viability was evaluated by MTT assay after 24­h. To show apoptosis inductionof breast and prostate­ cancer cell death by extracts, Annexin/PI test were performed. BCL₂ gene expression was analyzed using Real-Time RT-qPCRfor 24,48h.
Results: The highest effects of growth inhibition were observed in the prostate, breast and fibroblast cell lines with extract, respectively. The combined effect of doxorubicin with extract in three cell lines in comparison with control did not show any significant difference. The results of Annexin / PI indicated that the percentage of initial apoptosis, delayed  apoptosis and necrosis in treated cells increased compared to control. BCL₂ expression in cell lines decreased significantly over 24 and 48 h compares to control(p < 0.01).
Conclusion: hydroalcoholic extract of B.persica seed has ability to inhibit the proliferation of cancer cell lines as well as the induction of apoptosis in cancer cells, compared with using it with doxorubicin, although the extract did not have any synergy effect with doxorubicin at lower concentrations, it can be a substituted for this kind of drug with fewer side effects.
 

Abstract Aim: 7-bromoindirubin-3'-oxime (7-BIO) is a well-known glycogen synthase kinase-3β (GSK3β) inhibitor with anticancer effects on different cancer cell lines. However, due to its low water solubility, absorption and high gastrointestinal toxicity, it has been inapplicable in clinics so far. In the present study, bovine serum albumin nanoparticles (BSA NPs) were prepared as drug carriers.
 Material and methods: to be conjugated with 7-BIO to increase the effectiveness and reduce the toxicity. The nanoparticles were synthesized and conjugated with 7-BIO and their sizes were analysed by using scanning electron microscopy. Also, the viability and apoptosis in the cells treated with 7-BIO conjugated nanoparticles were examined by MTT assay and acridine orange/ethidium bromide staining (AO/EB), respectively. The significance of the data variation was evaluated by using ANOVA test.
Results: 10 mg/ml albumin led to spherical nanoparticles (NPs) of 89.42±7 nm in diameter with mono dispersity. MTT assay showed that 7Bio-loaded BSA NPs had a higher toxic effect on MCF-7 cell line compared to the free 7-BIO. Also, AO/EB staining showed that the apoptosis was induced by 7-BIO treatments. 7-BIO is known to specifically inhibit GSK3β phosphorylate GSK3β, which was also evident in our treated breast cancer cell line MCF-7 both with free and loaded BSA NPs 7Bio.
 Conclusion: The albumin nanoparticles conjugated with the GSK3 inhibitor effectively decreased the viability of the proliferating breast cancer cell line MCF-7, pointing at its possible clinical impact in future.
 
 

Abstract Aim: The aim of this study was to evaluate the effect of silymarin and lithium chloride on DNA integrity and nucleus of ram sperm.
Material and Methods: In this study, Farahani's ram testes were obtained from Arak slaughterhouse immediately after ram daily slaughter and transferred to the research laboratory. A few incisions were made in the epididymis, and spermatozoa were  then washed into a sterile falcon tube by Ham's F10 medium. collected spermatozoa of ram were divided into four groups: 1. Sperm at 0 hour, 2. Sperm incubated for180 minutes (control), 3. Sperm treated with lithium chloride for 180 minutes and 4. Sperm treated with silymarin + lithium chloride for 180 minutes. DNA integrity and DNA fragmentation were investigated by acridine orange staining sperm chromatin expersion (SCD) test respectively. Morphological feature of apoptosis in sperm nucleus was assessed using Diff-Quick staining. Data were analyzed using one-way analysis of variance test (ANOVA) followed by Turkey's test .
Results: The percentage of DNA fragment and apoptosis were significantly increased in lithium chloride-treated group compared to the control. In silymarin+ lithium chloride group, Silymarin could signicantly compensate these effect compared to the lithium choloride group.
Conclusion: Silymarin as a potent antioxidant could  prevent toxic effect of  lithium  on DNA  fragmentation and apoptosis in sperm nucleus.
 

Abstract Aim: The aim of this study is investigation of cell toxicity of oxaliplatin on colon cancer cell line and analysis of caspase 3 and 9 apoptotic genes.
Material and methods: In this experimental study, cytotoxicity of oxaliplatin on colon cancer cell line (HT29) was evaluated using MTT method in different concentrations including 100, 50, 25, 12.5, 6.25 and 3.125 µg/mL and the IC50 value was determined. After treatment of HT29 cells with IC50 value, the cells RNA was extracted and converted to cDNA. Subsequently, the expression level of caspase 3 and caspase 9 apoptotic genes comparing to house-keeping gene (β-actin) was measured using Real Time PCR.
Results: Treatment of HT29 cells with various concentrations of oxaliplatin including show that oxaliplatin has the highest cytotoxic effect in 100 µg/mL, which is statistically significant (p < 0.05). In addition to, the IC50 value of oxaliplatin was 6 µg/mL. Moreover, the expression ratio of caspase 3 and 9 genes comparing to β-actin gene in HT29 cells treated with oxaliplatin were up-regulated (2.69±0.72 (p < 0.001), 3.26±0.56 (p < 0.001), respectively).  
Conclusion: According to cell toxicity and apoptosis induction in HT29 cells by oxaliplatin, it can be concluded that this drug is an appropriate choice for treating of colon cancer.
 

Abstract Aim: In this study, the effects of simulated microgravity on the apoptosis and expression of p75NTR gene were investigated in the adipose derived stem cells before and after the neural differentiation.
Material and Methods: Human adipose derived stem cells were isolated, cultured and differentiated.  A single-axis clinostat apparatus was used to simulate microgravity for 6, 24 and 72 hours. Real time PCR technique was used for gene expression analysis after extraction of RNA of samples. Cell viability was assessed by MTT assay and apoptosis rate was calculated by Annexin V staining.
Results: Our results showed that microgravity led to a significant decrease in p75NTR gene expression in adipose derived stem cells. However, microgravity had no significant effect on viability of cells before and after differentiation, but apoptosis in undifferentiated cells was decreased in contrast to controls.
Conclusion: Due to reduction of apoptosis and increment of differentiation potential of cells, microgravity can be introduced as a powerful tool and also new condition for cell culture and neural differentiation for achievement to effective cell therapy. 
 

Abstract Aim: The aim of this study is evaluating the effect of crocin as one of saffron bioactive compounds in mice spermatogenic stem cells.
Material and Methods: Balb/c neonate spermatogenic stem cells were grown in DMEM-F12 medium and were treated with various concentrations of crocin (2.5, 5, 10, 20, 40 µg/ml) for 6 and 12 days. For detecting spermatogenic stem cells, assessment cytotoxicity, recognizing viable cells and antioxidant capacity have been used alkaline phosphatase assay, MTT assay, AO, DAPI staining, and DCF-DA assay, respectively. Statistical analyses, One-way ANOVA, Duncan test(P≤0.05), were conducted using SPSS software .
Results: Crocin in concentrations >20 µg/ml exerted no significant toxicity on mice spermatogenic stem cells in 6 and 12 days treatment. Meanwhile, viability of mice spermatogenic stem cells was reduced under exposure to 20, 40 µg/ml to 21, 24 % in 6 days and 29, 41% in 12 days treatment. Further, evaluations of cell viability by fluorescence microscopy and antioxidant potential by fluorimetery have showed protective and antioxidant effect of crocin after 12 day treatment.
Conclusion: Crocin can be induced protective effect on mice spermatogenic stem cells, which conducted their effect through minimal effects on cell viability and reducing cell death.

Abstract Aim: In this study, the effects of curcumin- coated silver nanoparticles were examined on induction of apoptosis in ovarian cancer A2780 cell.
Material and Methods: Silver nanoparticles coated with curcumin were biosynthesized, then A2780 cells were treated with different concentration of silver nanoparticles. Cytotoxicity effects of silver nanoparticles in A2780 cells were assessed by MTT assay and apoptotic effects of these nanoparticles were examined using DAPI and acridine orange/ propidium iodide staining and caspase 3/9 activation assay. Changes in Bax and Bcl-2 gene expression were analyzed by Real Time PCR.
Results: Findings showed that silver nanoparticles inhibit A2780 cells proliferation in a dose dependent manner. 8 µg/ml 50% decreased cell viability at 24 hours, DAPI and, acridine orange propidium iodide staining represent that percentage of apoptotic cells in the treated groups was increased. The results of Real Time PCR showed that Bax gene expression in cells treated with silver nanoparticles increased, while Bcl-2 gene expression in the treated cells was decreased.
Conclusion: Silver nanoparticles coated with curcumin induce apoptosis in A2780 cancer cells. The use of the nanoparticles should be considered a promising strategy for the treatment of ovarian cancer.

Abstract Aim: This study aimed to investigate the effect of Bisphenol­ A on the cell viability, morphologic changes and induction of apoptosis in bone marrow mesenchymal stem cells of an adult rat. Material and Methods: The bone marrow mesenchymal stem cells were extracted using flashing-out method. At the end of the third passage, cells were divided into 11 groups of control and experimental. Experimental cells were treated with the different doses of Bisphenol­ A (1, 5, 10, 50, 100, 250, 500, 1000, 2000 and 4000 nM) for four periods of 5, 10, 15 and 21 days in the osteogenic media containing 10% of fetal bovine serum. The cell viability, DNA damage, expression of genes and the morphologic changes of the cells were then investigated during the procedure of osteogenesis. The study data was analyzed using one-way ANOVA and T-Test setting the significant P value at p < strong>Results: Within Bisphenol­ A treated cells, the mean viability, the mean of nuclei diameter and the expression of anti-apoptotic Bcl-2 gene of the mesenchymal stem cells treated with Bisphenol­ A significantly decreased (p < 0.05), compared to the control group. In addition the DNA damage and the expression of apoptotic Bax gene of the cells treated with Bisphenol­ A significantly increased, compared to the control group (p < 0.05). Conclusion: Our data suggest that BPA decreases cell viability and induces apoptosis in mesenchymal stem cells, in a dose dependant manner.

Abstract Aim: The aim of this study was to assess side effects of clinical dose of busulfan on testis and epididymal sperm of adult male mouse. Material and Methods: Male adult NMRI mice (25-35 g) were divided into two groups of sixteen each. Control and busulfan treated group were administered 100 μL DMSO (Dimethyl Sulfoxide) and 3.2 mg/kg busulfan for 4 days, respectively. The animals were sacrificed 35 days after starting treatment. The histological change on germinal epithelium, sperm quality parameters (count and normal morphology), viability and DNA fragmentation on sperm were analyzed by light microscopy, CASA (Computer–aided Sperm Analyzer), MTT (3-[4, 5-Dimethylthiazol-2-yl]-2, 5 Diphenyltetrazolium Bromide) and TUNEL assays, respectively. Results: Busulfan administration significantly decreased parameters of sperm (count and normal morphology) and increased germinal epithelium destruction. The head, mid piece and tail abnormalities of treated group were increased significantly versus control. The lower levels of MTT in treated group were significant compared with control group. Higher levels of TUNEL positive cells that were found in treated groups demonstrated the increasing of DNA fragmentation in sperms following busulfan treatment. Conclusion: The results showed that clinical dose of busufan induces genotoxic, pre-apoptotic and cytotoxic effects on spermatogenesis. Also, busulfan can increase cytotoxicity on testis tissue of adult male mouse.