Cell and Tissue Journal

Abstract Aim: Breast cancer remains a significant health challenge, being one of the most common cancers among women and a leading cause of cancer-related deaths globally. The need for effective treatment options is paramount, and recent research has spotlighted sulfonamide derivatives for their notable antitumor properties. These compounds have shown considerable cytotoxic effects against various cancer cell lines, including breast cancer, through mechanisms such as apoptosis induction and inhibition of heat shock protein 70 (HSP70), which is crucial for cancer cell survival. This study focuses on synthesizing a solid lipid nanocarrier that incorporates a sulfonamide derivative, aiming to enhance its anticancer efficacy against the MCF-7 breast cancer cell line. The encapsulation of sulfonamide within these nanoparticles is expected to improve its bioavailability by increasing solubility and stability, thereby allowing higher concentrations to effectively reach tumor sites. Additionally, these nanoparticles are designed to minimize systemic toxicity and protect healthy tissues from adverse effects, resulting in enhanced therapeutic efficacy.
methods: The research involved synthesizing solid lipid nanoparticles (SLNs) containing the sulfonamide derivative, followed by a comprehensive analysis of their physicochemical properties. Key parameters assessed included drug loading capacity, morphology, size distribution, and release profile using techniques such as Fourier Transform Infrared Spectroscopy (FTIR), Dynamic Light Scattering (DLS), Atomic Force Microscopy (AFM), and dialysis methods. The cytotoxicity of the nanocarrier was evaluated on MCF-7 cells through the MTT assay, alongside assessments of cell proliferation and migration using scratch assays.
Results: The synthesized nanocarrier had an average size of 315 nanometers with a Polydispersity Index (PDI) of 0.3, indicating good uniformity. AFM analysis confirmed its spherical morphology. The nanocarrier demonstrated a high drug loading capacity of approximately 82% for the sulfonamide derivative and facilitated a controlled release over time. Cytotoxicity assays revealed that while the blank nanocarrier was safe, the formulation containing the sulfonamide derivative exhibited significantly enhanced cytotoxic effects on MCF-7 cells at lower concentrations compared to the free sulfonamide alone. This improvement in efficacy is attributed to the controlled release mechanism that allows for sustained exposure of cancer cells to the active compound. Moreover, the nanocarrier was more effective in inhibiting both growth and migration of cancer cells than treatments with free sulfonamide, as evidenced by scratch assays.
Conclusion: This research highlights the potential of solid lipid nanoparticles in improving the therapeutic effectiveness of sulfonamide derivatives against breast cancer. The synthesized nanocarrier demonstrates favorable physicochemical properties, including high drug loading capacity and controlled release, which are essential for effective treatment outcomes. Notably, encapsulating sulfonamide derivatives significantly enhances cytotoxicity against MCF-7 cells at lower doses compared to their unencapsulated counterparts, emphasizing its promise for improved therapeutic strategies. Furthermore, these nanoparticles effectively inhibit cell proliferation and migration, contributing to tumor growth suppression. This innovative approach aims not only to enhance treatment efficacy but also to minimize systemic toxicity and protect healthy tissues from adverse effects. Future research should focus on innovative clinical applications and further optimization of these nanocarriers to maximize their potential in oncology, paving the way for more effective treatments for breast cancer.

Abstract Aim: Cancer is one of the most important diseases of this century, which affects many people all over the world, and while the prevalence of this disease is increasing, no suitable treatment has been found yet. In the meantime, breast cancer is the most common cancer and the leading cause of cancer-related death in women worldwide. In the search for cancer treatment solutions, researchers synthesize numerous medicinal compounds for various cancers and study their effects. Sorafenib is a urea multikinase inhibitor which causes apoptosis and inhibits angiogenesis and cancer cell proliferation. This compound is currently used as a treatment for hepatocellular cancer, and tests have also been carried out to verify its performance in treating other cancers. Sorafenib derivatives were also synthesized and analyzed. In this research, the anticancer effect of one of the derivatives of sorafenib named (2E,2´E)-2,2´-(1,4-phenylene bis(methanylidene)bis(N-(4-chloro-3-(tri) Fluoroethyl (phenyl) hydrazine carboxamide, abbreviated as SO-D-3, was evaluated against three breast cancer cell lines.
Materials and methods: Three breast cancer cell lines including MCF7, 4T1 and MDA were prepared and cultured in RMPI culture medium. Cells were treated with concentrations of 31.25, 62.5, 125, 250, 500, 1000 and 2000 μM of SO-D-3, and their survival rates were measured using the MTT test during 24, 48 and 72 hours. The induction of apoptosis by SO-D-3 in the studied cancer cells was investigated by flow cytometry. In addition, the level of Hsp70 and Casp8 proteins as implicated in apoptosis were investigated using the western blot method. The results were statistically analyzed using SPSS software.
Results: The results of the MTT test showed that the treatment of cancer cells significantly decreases the survival of these cells compared to the control. The IC50 for MDA-MB-231 cells was significantly lower than MCF7 and 4T1 cells at 24 hours (157.4 µM for MDA-MB-231 compared to 843.8 µM for MCF7 and 1212 µM for 4T1). However, the difference decreased with increasing treatment time (77.47 μM and 23.38 μM for MDA-MB-231, 107.3 μM and 36.69 μM for MCF7, and 83.63 μM and 16.58 μM for 4T1 at 48 hours and 72 hours respectively). Flow cytometry analysis revealed that in cells treated with SO-D-3, apoptotic cells increased significantly. Examination of the protein level using the western blot technique also showed that the HSP70 protein was significantly decreased in MCF7 and 4T1 cells. On the other hand, the level of Casp8 protein was significantly increased in 4T1 and MDA cells.
Conclusion: The results of the present study showed that SO-D-3 can cause the death of breast cancer cells in a time-dependent manner. Subsequent experiments showed that SO-D-3 exerts this action through the induction of apoptosis. Therefore, according to the results of the present research, SO-D-3 can be introduced as a compound with potential anti-cancer properties that can cause cell death by reducing Hsp70 and changing the expression of Casp8 involved in the external pathway of apoptosis.

Abstract

Abstract Aim: The role of Hsp70 chaperone from Rutilus frisii kutum in the thermal inactivation of luciferase in E. coli cell carrying Hsp70 and firefly luciferase was investigated.
Material and Methods: Co-transformation of E. coli cell was carried out with two expression vectors containing Hsp70 and firefly luciferase. The co-transformed cells carrying Hsp70 and luciferase were expressed under optimum conditions. After adding tetracycline, the cells were then incubated for 60 min at 40, 42, 44, 46, 48 and 50°C treatments. Finally, the luminescence activity of samples was calculated.
Results: After 30 min at 40 and 42°C temperatures, the luciferase activity in control samples reached almost zero, while in the co-transformed samples, about 72% and 60% of the luminance activity maintained compared with control samples (before heat treatment), and even after 60 min, up to 36% and 22% of the activity remained. Also, at 44 and 46°C temperatures in the initial times after stress, a significant difference was observed between the luciferase activity of the co-transformed and the control samples. In contrast, at 48 and 50°C temperatures, the changes of the luciferase activity of co-transformed samples were small compared with control samples even in the early stages of stress.
Conclusion: The thermal aggregation of luciferase as a significant reporter protein is inhibited at high temperatures via the activity of Hsp70 in the bacterial cell, which this process can be widely used in the food and pharmaceutical industries and the providing cancer diagnostic kits.
 

Abstract Aim: The aim of the present study was to investigate the optimal expression of Hsp70 recombinant Rutilus FrissiKutum liver in E. coli.
Material and methods: To evaluating of expression level of recombinant Hsp70, transformed E. coli were cultured in LB medium at 2, 4, 8, 12, 24, 36 and 48 hours, 20, 25, 30, 37 and 50 °C, extreme pH (5 and 9) and in present of heavy metals e.g. mercury, cobalt, iron, zinc and etc. Expression of recombinant Hsp70 protein was determined by SDS-PAGE 12% analysis of E. coli extracts followed by staining with Coomassie Blue.
Results: Optimum expression of recombinant Hsp70 was obtained at 4 and 8 h, 37 °C and pH 5. Additionally, the maximum and minimum expression of this protein was achieved in present of manganese and cobalt respectively. The results further indicate that the survival of E. coli with Hsp70 was enhanced compared to the control cells.
Conclusion: All results reveal that transformed bacteria are able to survive against extreme environmental conditions compared to control sample, because of having recombinant Hsp70. This heat-shock protein protects vital proteins from denaturing by its chaperone property. So, growth condition described in this study can be used for optimizing production of recombinant Hsp70 in E. coli host.