In vitro analysis of single chain variable fragment-based immunotoxins against Erythropoietin-producing hepatocellular A2 receptor overexpressed in breast cancer cells.

Breast cancer is one of the leading causes of cancer deaths worldwide. Thereafter, designing new treatments with higher specificity and efficacy is urgently required. In this regard, targeted immunotherapy using immunotoxins has shown great promise in treating cancer. To target a breast cancer cell, the authors used the antibody fragment against a receptor tyrosine kinase, EphA2, which is overexpressed in many cancers. This fragment was conjugated to a plant toxin, subunit A of ricin, in two different orientations from N to C-terminal (EphA2- C-Ricin and EphA2- N-Ricin). Then, these two immunotoxins were characterized using in vitro cell-based assays. Three different cell lines were treated, MDA-MB-231 (breast cancer) which has a high level of EphA2 expression, MCF-7 (breast cancer) which has a low level of EphA2 expression, and HEK293 (human embryonic kidney) which has a very low level of EphA2 expression. Moreover, binding ability, cytotoxicity, internalization, and apoptosis capacity of these two newly developed immunotoxins were investigated. The flow cytometry using Annexin V- Propidium iodide (PI) method indicated significant induction of apoptosis only in the MDA-MB-231 cells at different concentrations. It was also found that construct I, EphA2- C-Ricin immunotoxin, could bind, internalize, and induce apoptosis better than the EphA2- N-Ricin immunotoxin. In addition, the obtained data suggested that the N or C-terminal orientation conformation is of significant importance.

Analyzing of colorectal cancerrelated genes and microRNAs expression profiles in response to probiotics Lactobacillus acidophilus and Saccharomyces cerevisiae in colon cancer cell lines.

BACKGROUND: Colorectal cancer is the world's third most frequent cancer and the fourth cause of mortality. Probiotics play an important function in preventing metastasis as well as the growth and proliferation of malignant cancer cells. METHODS AND RESULTS: The study investigated the anticancer effect of Lactobacillus acidophilus supernatant and Saccharomyces cerevisiae yeast on colorectal cell lines, including HT29 and SW480 as a colorectal cancer model. The extract from the Lactobacillus acidophilus and Saccharomyces cerevisiae standard probiotics were prepared, and probiotics characterization was confirmed by morphological and Biochemical tests. The viability of HT29 and SW480 colon cancer cell lines on effecting probiotic supernatant was evaluated by measuring the MTT colorimetric method. Comparison of the expression profile of several genes involved in apoptosis, cell cycle, and metastatic pathway in HT29 and SW480 cell lines with the treatment of probiotics extract showed an upregulation in the BAX, CASP3, and CASP9 and down regulation BCl-2, MMP2, and MMP9 genes. Also, a comparison of microRNA expression profiles indicated an increase of miR 34, 135, 25, 16, 195, 27, 98, let7 and a decrease of miR 9, 106b, 17, 21, 155, 221. CONCLUSIONS AND DISCUSSION: The findings of this study indicate that probiotics can effectively suppress the proliferation of colorectal cancer cells and even reverse their development. Additionally, the study of cellular genes and miRNA profiles associated with colorectal cancer have demonstrated that our probiotics play a crucial role in CRC prevention by increasing the expression of tumor suppressor microRNAs and their target genes while decreasing oncogenes.

Hydroxyurea-loaded Fe3O4/SiO2/chitosan-g-mPEG2000 nanoparticles; pH-dependent drug release and evaluation of cell cycle arrest and altering p53 and lincRNA-p21 genes expression.

Carbohydrate polymers were widely used in pharmaceuticals and drug delivery systems due to their biodegradability and biocompatibility. Among them, chitosan (Cs) has been considered in many new drug delivery systems. Poly(ethylene glycol) as a hydrophilic polymer can increase the solubility and stealth functions of nanocarriers. The Fe3O4 nanoparticles functionalized with polymers act as non-toxic drug vehicles for tumor targeting under external magnetic fields. In present study, the Fe3O4/SiO2-NH2 nanoparticles were prepared and then functionalized with methoxy-PEGylated chitosan (Cs-g-mPEG2000) and the hydroxyurea (HU) was loaded on this nanoparticles. The structure, crystallinity, and morphology of HU/Fe3O4/SiO2/Cs-g-mPEG2000 were determined using spectroscopic and electron microscopy analysis. Encapsulation efficiency of HU and the percentage of loading and release rate at different pH values at 37 °C were examined. Maximum drug release was observed at pH = 7.4. According to TEM results, the nanoparticle sizes were between 18 and 157 nm. The cytotoxicity effect of HU-loaded nanoparticles against MCF-7 human breast cancer cell was evaluated using MTT assay and cell cycle arrest analysis. The inhibitory concentration (IC50) values were 249 and 85 µg/mL on the MCF-7 cell line compared to the control group in 24 h and 96 h, respectively. In addition, the expression of p53 and lincRNA-P21 genes in treated cells and control group was assessed using real-time PCR, and the results showed that the ratio of p53 expression to lincRNA-P21 in MCF-7 cells was significantly increased (P < 0.05). The cell cycle arrested in the S-phase and the population of cells increased 1.3-fold compared to the control group.

Fluorescent tamoxifen-encapsulated nanocapsules functionalized with folic acid for enhanced drug delivery toward breast cancer cell line MCF-7 and cancer cell imaging.

Nanoscale drug delivery systems such as nanocapsules at the convergence of nanotechnology and biomedical sciences have been widely used. In the present study, with the aim of simultaneous imaging and therapy of cancer cells based on biodegradable/biocompatible polymers, we designed and synthesized tamoxifen-encapsulated nanocapsules to target the folate receptor positive breast cancer cells. Noteworthy, to monitor and link to the cancer cells, these nanocapsules were functionalized with fluorescein isothiocyanate and folic acid. The synthesized nanocapsules were characterized by FTIR, XRD, and PL spectroscopy, as well as FESEM and TEM techniques. Although the free tamoxifen has low solubility in physiological solutions, the synthesized tamoxifen-encapsulated nanocapsules have enough solubility, good stability, and more biocompatibility in these solutions. The encapsulation of tamoxifen into the nanocapsules, tamoxifen loading, and its subsequent release behavior were studied. In order to investigate the biological role of these nanocapsules, MTT assay and cell imaging analysis have also been examined. The cytotoxicity test exhibit that the mean IC50 values on the MCF-7 cell line were found to be 15.52 and 8.46 µg/ml in 24 h and 48 h respectively and the cytotoxicity increased by approximately 2.72-fold compared with free TAM against the MCF-7 cancer cell line. Also, cell imaging experiments showed that the synthesized nanocapsules have appropriate cellular uptake efficiency, good potential for monitoring of these particles in vitro. The experimental results suggest that the synthesized tamoxifen nanocapsules facilitate the proper targeting, drug encapsulation efficiency, and controlled release of tamoxifen in vitro.

Silver nanoparticles biosynthesised using Centella asiatica leaf extract: apoptosis induction in MCF-7 breast cancer cell line.

The aim of this study was to green synthesised silver nanoparticles (AgNPs) using Centella asiatica leaf extract and investigate the cytotoxic and apoptosis-inducing effects of these nanoparticles in MCF-7 breast cancer cell line. The characteristics and morphology of the green synthesised AgNPs were evaluated using transmission electron microscopy, scanning electron microscopy, UV-visible spectroscopy, X-ray diffraction, and Fourier-transform infrared spectroscopy. The MTT assay was used to investigate the anti-proliferative activity of biosynthesised nanoparticles in MCF-7 cells. Apoptosis test was performed using flow cytometry and expression of caspase 3 and 9 genes. The spherical AgNPs with an average size of 19.17 nm were synthesised. The results showed that biosynthesised AgNPs exhibited cytotoxicity, anti-cancer, apoptosis induction, and increased expression of genes encoding for caspases 3 and 9 in MCF-7 cancer cells in a concentration- and time-dependent manner. It seems that green synthesised AgNPs have potential uses for pharmaceutical industries.