Cell and Tissue Journal

Abstract Introduction: Prostate cancer is one of the most prevalent malignancies among men worldwide and represents a major cause of cancer related morbidity and mortality. Despite advances in screening and therapeutic approaches, resistance to standard treatments, including androgen‑deprivation therapy, remains a major clinical challenge in prostate cancer. As a result, identifying novel compounds that can suppress proliferation or modulate key molecular regulators of the cell cycle is of considerable interest. Papaverine is an isoquinoline alkaloid derived from the opium poppy, widely known for its long‑standing clinical use as a smooth‑muscle relaxant and vasodilator. In recent years, it has gained attention for its potential anticancer effects, including its influence on mitochondrial function, cellular energy metabolism, and the modulation of signaling pathways involved in cell cycle regulation. However, the specific effects of papaverine on prostate cancer cells have not yet been clearly elucidated. Tamoxifen, a selective estrogen receptor modulator widely used in breast cancer therapy, has also demonstrated off target antiproliferative effects in various tumor models.
Aims: The present study aimed to evaluate the effects of papaverine and tamoxifen on the viability and molecular regulatory profile of DU145 prostate cancer cells line. Specifically, the study investigated the cytotoxic potential of these compounds on different concentrations and incubation times, and their ability to alter the expression of CDK4 gene, a key mediator of G1 to S phase progression, along with two microRNAs, miR-146a and miR-22, known to influence cell cycle control and tumorigenic pathways. By integrating cellular and molecular findings, the study sought to clarify whether these compounds could serve as potential modulators of prostate cancer cell growth.
Materials and Methods: DU145 human prostate cancer cell line were cultured under standard conditions and treated with different concentrations of papaverine and tamoxifen. Cell viability was assessed for 24, 48, and 72 hours using the MTT assay (3 [4,5 dimethylimidazole-2-yl] 2,5 diphenyl tetrazolium bromide). Dose–response curves were generated to calculate IC50 values for each compound at the specified time points. To investigate molecular changes, total RNA was extracted from the treated and the control cells, and the expression levels of CDK4 gene, miR-146a, and miR-22 were quantified using Real Time PCR with appropriate internal controls. Relative expression changes were determined using the 2-ΔΔCt method. The data were analyzed using SPSS software. Results were presented as Mean ± SD, and significance level of p ≤ 0.05 was considered statistically significant.
Results: Treatment of DU145 prostate cancer cell line with various concentrations of papaverine and tamoxifen resulted in a significant reduction in cell viability in a concentration and time dependent manner, as measured by the MTT assay for 24, 48, and 72 hours. IC50 values were successfully determined for both compounds across the different time points.
Molecular analysis using Real Time PCR showed that treatment with either papaverine or tamoxifen led to a marked downregulation of CDK4 expression compared with untreated controls. In contrast, both compounds induced a significant up‑regulation of miR‑146a and miR‑22 levels.
Discussion: The observed decrease in viability suggests that papaverine and tamoxifen exert notable antiproliferative effects on DU145 prostate cancer cells. The downregulation of CDK4 gene provides a mechanistic link to cell cycle arrest, as CDK4 is a central regulator of G1‑to‑S phase progression. Such suppression is in line with reduced cellular proliferation and supports the therapeutic potential of these compounds.
The increased expression of miR‑146a and miR‑22 further highlights the involvement of tumor suppressive microRNAs in mediating these effects. Both miRNAs have been implicated in regulating pathways associated with proliferation, apoptosis, and oncogenic signaling. Their upregulation may contribute to the inhibition of cell growth through negative modulation of critical oncogenic targets.
These results align with prior evidence demonstrating tamoxifen’s antiproliferative effects but also importantly emphasize that the molecular and cellular impacts of papaverine in prostate cancer remain poorly understood. The current findings provide initial insight into papaverine’s interaction with cell cycle regulatory genes and microRNAs, suggesting a potential role in prostate cancer growth inhibition. Considering the adverse effect profile associated with tamoxifen, papaverine may therefore represent a more favorable candidate for further investigation, particularly as a potential alternative with a potentially safer therapeutic window.

Abstract Aim: In order to study the efficiency of the tuber specific promoter (Patatin1), the expression of the HBsAg antigen of the hepatitis B vaccine was evaluated using a transient expression method (AgroInfiltration) in the tubers and leaves of potato plants.
Material and Methods: A tuber specific promoter (Pat1) was isolated from the potato plant using a specific primer pairs by Polymerase Chain Reaction (PCR). It was cloned in the upstream of a synthetic optimized codon of the HBsAg gene in the binary plant vector pBI121. In order to compare the tissue specificity of Pat1, the HBsAg gene also was used under the control of a consultative CaMV35S promoter. The genetic constructs were transferred to the tubers and leaves of potato plants using the Agroinfiltration method. An HBsAg antigen content was measured using ELISA method.
Results: The level of HBsAg antigens in the leaves and tubers of potato plants indicated that Pat1 promoter specifically induced HBsAg high expression in the tuber tissues. Very low expression by the Pat1 promoter in the leaf tissues has been also reported and can be partly dependent on the presence of inducing factors in the inoculation media. However, the expression of HBsAg antigen occurs in both leaf and tuber tissues under the consultative promoter CaMV35S.  The results showed that the codon optimized HBsAg gene for potato plant was expressed properly in plant tissues.
Conclusion:  findings indicate that potato tuber specific promoter Pat1 can be used effectively to express the synthetic optimized HBsAg antigen in the transgenic potato plants.
 
 

Abstract Aim: The current research was done to investigate the sperm plasma membrane integrity, oxidative stress factors of sperm and testis in male mice treated with cadmium.
Material and Methods: 24 adult NMRI mice were divided into four groups: 1. Control group 2.  Treated with cadmium chloride (5 mg/kg) 3. Treated with Curcumin (100 mg/kg) 4. Treated with curcumin + cadmium chloride. The treatments were performed as a single dose and after 24 hours, epididymal spermatozoa of different groups were used to evaluate the plasma membrane integrity. In addition, the amount of malondialdehyde (MDA) and the anti-oxidant activity of serum and testis were evaluated. Data were analyzed using ANOVA test followed by Turkey’s test (p < 0.05).
Results: findings showed that cadmium chloride caused a significant decrement in plasma membrane integrity compared to the control groups. In addition, cadmium chloride induced a significant increment of MDA in serum and testis and a significant decrement in total antioxidant activity of serum and testis as compared to the group. In curcumin + cadmium chloride- treated group, curcumin could significantly compensate the toxic effect of cadmium on these parameters compared to the cadmium-treated group.
Conclusion: it seems thatcurcumin as a potent antioxidant is able to ameliorate the adverse effects of cadmium chloride on sperm plasma membrane integrity, lipid oxidation and total antioxidant activity of serum and testis.
 

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.