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Abstract This study highlights the cytotoxic effect of three L. casei strains on colorectal cell lines in invitro conditions. Different concentrations of live, heat killed (HK) and cell free supernatant (CFS) of three L.casei strains were subjected to CaCo2 and MRC5 cell lines. The viability of the treated and untreated cells was determined after 72 hrs by MTT assay, and IC50 estimated. Apoptosis was evaluated by Annexin V-propidium iodide method using flow cytometry. The live, HK and CFS of the L. casei strains showed cytotoxic effects on colorectal cell lines with significant differences. The cytotoxicity effects of live cells on CaCo2 cells were significantly higher (p˂0.01) than the HK cells. A dose dependent response was observed, as higher concentrations resulted in enhanced cytotoxicity effects. Live L.casei 1296-2cells inhibited 91% of CaCo2 cell growth, with IC50 of less than 108 cfu/ml. MRS medium and concentrations of CFS at above 20% v/v, were cytotoxic to the normal cell lines. Flow cytometry analyses of L. casei 1296-2 indicated that cytotoxicity effects on CaCo2 cells is related to apoptotic induction. Invitro studies indicate that Live and CFS of L. casei 1296-2 might be promising candidate for the control of colorectal cancers.

Exosomes, a small bilayer membrane derived from eukaryotic cells, have been identified as a useful natural delivery platform due to their suitable size, biocompatibility, structural stability, high loading capacity, and editable surface capability. Due to the difficulty of maintaining the highly pure exosome, several attempts have conducted the techniques for exosome isolation. In recent years, microstructures have found many applications in chemistry, biology, and medicine due to their high accuracy and low cost of materials. Soft lithography is a low-cost, fast, accurate, and yet widely used method of construction of Micron channels. In the present study, a soft lithography process has been performed to construct channels for exosome separation with immunoaffinity function. Both biochemical and biophysical categories tests were performed to examine the quality of extracted exosomes from different sources (serum, cell supernatant, and urine) and compared with the commercially available kit. Results showed that the current technique was capable to isolate exosomes with a high yield rate, purity, and low time consumption. All forms of the imatinib loaded exosomes exhibited the antitumor activity against KYO-1 cell line. [ABSTRACT FROM AUTHOR]

Leukemia, a type of cancer that affects the blood and bone marrow, is a complex and devastating disease with limited treatment options. Over the years, researchers have been exploring novel therapeutic approaches to improve patient outcomes. One emerging area of interest is the role of exosomes in leukemia. Exosomes are small extracellular vesicles secreted by various cell types, including cancer cells. They play a crucial role in intercellular communication by transferring bioactive molecules such as microRNAs (miR- NAs) between cells. Recent studies have highlighted the involvement of exosome miRNAs in leukemia progression, therapy resistance, and disease prognosis. Understanding the mechanisms underlying exosome-mediated miRNA transfer in leukemia may provide valuable insights into disease pathogenesis and potential therapeutic targets. Furthermore, exosomes can serve as non-invasive biomarkers for leukemia diagnosis and monitoring treatment response. Studies have demonstrated that high levels of galectin-3 are associated with worse clinical outcomes in patients with leukemia. Understanding the specific mechanisms by which galectin-3 contributes to leukemogenesis could provide valuable insights into novel therapeutic strategies for this devastating disease. We characterized the abundance of exosomes as a function of the number and distribution in the samples. 48 h post downregulation of the LGALS3 gene, we observed a 1.2-fold increase in the release of exosomes compared to the control. [ABSTRACT FROM AUTHOR]

Purpose: Fetal hemoglobin (HbF) upregulation is a mitigating factor in ß-hemoglobinopathies therapy like ß-thalassemia and sickle cell diseases. Finding molecular mechanisms and the key regulators responsible for globin switching could be helpful to develop effective ways to HbF upregulation. In our prior in silico report, we identified a few factors that are likely to be responsible for globin switching. The goal of this study is to experimentally validate the factors. Methods: We established K562 cell line with BCL11A knock down leading to increase in HBG1/2 using CRISPR/Cas9 system. Then, using quantitative polymerase chain reaction (qPCR), we determined the expression level of the factors which were previously identified in our prior in silico study. Results: our analysis showed that BCL11A was substantially knocked down, resulting in the upregulation of HBG1/2 in the BCL11A-ablated K562 cells using CRISPR/Cas9 system. Additionally, the experimental data acquired in this study validated our prior bioinformatics findings about three potentially responsible genes for globin switching, namely HIST1H2Bl, TRIM58, and Al133243.2. Conclusion: BCL11A is a promising candidate for the treatment of ß-hemoglobinopathies, with high HbF reactivation. In addition, HIST1H2BL, TRIM58 and Al133243.2 are likely to be involved in the mechanism of hemoglobin switching. To further validate the selected genes, more experimental in vivo and in vitro studies are required. [ABSTRACT FROM AUTHOR]

Objective Lactobacilli are a group of probiotics with beneficial effects on prevention of cancer. However, there is scant data in relation with the impacts of probiotics in late-stage cancer progration, especially metastasis. The present original work was aimed to evaluate the anti-metastatic and anti-proliferative activity of lactobacillus rhamnosus supernatant (LRS) and lactobacillus crispatus supernatant (LCS) on the human cervical and colon adenocarcinoma cell lines (HeLa and HT-29, respectively). Materials and Methods In this experimental study, the anti-proliferative activities of LRS and LCS were determined through MTT assay. MRC-5 was used as a normal cell line. Expression analysis of CASP3, MMP2, MMP9, TIMP1 and TIMP2 genes was performed by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR), following the cell synchronization. Results Supernatants of these two lactobacilli had cytotoxic effect on HeLa, however LRS treatment was only effective on HT-29 cell line. In addition, LRS had no side-effect on normal cells. It was shown that CASP3 gene expression has been reduced after treatment with supernatants of two studied lactobacilli. According to our study, LRS and LCS are efficacious in the prevention of metastasis potency in HeLa cells with decreased expression of MMP2, MMP9 and increased expression of their inhibitors. In the case of HT-29 cells, only LRS showed this effect. Conclusion Herein, we have demonstrated two probiotics which have anti-metastatic effects on malignant cells and they can be administrated to postpone late-stage of cancer disease. LRS and LCS are effective on HeLa cell lines while only the effect of LRS is significant on HT-29, through cytotoxic and anti-metastatic mechanisms. Further assessments are required to evaluate our results on the other cancer cell lines, in advance to use these probiotics in other extensive trial studies.