Biogenic copper oxide nanoparticles from Bacillus coagulans induced reactive oxygen species generation and apoptotic and anti-metastatic activities in breast cancer cells.

The present study examined the anticancer capabilities of Bacillus coagulans supernatant-produced copper oxide nanoparticles (BC-CuONPs) on MCF-7 and SKBR3 cancer cells. The X-ray diffraction, ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, transmission electron microscopy, field-emission scanning electron microscopy, energy-dispersive X-ray, dynamic light scattering, and zeta potential techniques were used to characterize BC-CuONPs. This study also investigated the cellular and molecular processes of NPs' anti-proliferative and apoptotic properties on human breast cancer cells and compared them to the commercial pharmaceutical tamoxifen. The size of the spherical NP was from 5 to 47 nm with negative zeta potential. The MTT results showed the great cytotoxic effect of BC-CuONPs against breast cancer cells. The BC-CuONPs inhibited the growth of breast cancer cells in a time- and dose-dependent manner. The up-regulation of BCL2-associated X (BAX), cyclin dependent kinase inhibitor 1A (P21), Caspase 3 (CASP3), and Caspase 9 (CASP9), the down-regulation of BCL2 apoptosis regulator (BCL2), Annexin V-FITC/propidium iodide, and reactive oxygen species (ROS) generation results suggested that BC-CuONPs had a significant apoptotic impact when compared to the control. Scratch tests and vascular endothelial growth factor receptor gene (VEGF) down-regulation demonstrated that BC-CuONPs had anti-metastatic activity. The cell cycle analysis and down-regulation of Cyclin D1 (CCND1) and cyclin dependent kinase 4 (CDK4) revealed that cancer cells were arrested in the sub-G1 phase. Finally, the results showed that the secondary metabolites in the supernatant of Bacillus coagulans could form CuONPs, and biogenic BC-CuONPs showed anti-metastasis and anticancer properties on breast cancer cells while having less adverse effects on normal cells. Therefore, the synthesized CuONPs using B. coagulans supernatant can be shown as a potential candidate for a new therapeutic strategy in cancer management.

Exosomes of Mesenchymal Stem Cells as a Proper Vehicle for Transfecting miR-145 into the Breast Cancer Cell Line and Its Effect on Metastasis.

BACKGROUND: Despite recent advances in scientific knowledge and clinical practice, management, and treatment of breast cancer, as one of the leading causes of female mortality, breast cancer remains a major burden. Recently, methods employing stem cells and their derivatives, i.e., exosomes, in gene-based therapies hold great promise. Since these natural nanovesicles are able to transmit crucial cellular information which can be engineered to have robust delivery and targeting capacity, they are considered one of the modes of intercellular communication. miR-145, one of the downregulated microRNAs (miRNAs) in various cancers, can regulate tumor cell invasion, metastasis, apoptosis, and proliferation and stem cell differentiation. OBJECTIVES: The aim of this study was to investigate the role of exosomes secreted from adipose tissue-derived mesenchymal stem cells (MSCs) for miR-145 transfection into breast cancer cells in order to weaken their expansion and metastasis. METHODS: Here, we exploited the exosomes from adipose tissue-derived mesenchymal stem cells (MSC-Exo) to deliver miR-145 in the T-47D breast cancer cell line. Lentiviral vectors of miR-145-pLenti-III-enhanced green fluorescent protein (eGFP) and empty pLenti-III-eGFP as the backbone were used to transfect MSCs and T-47D cells. In order to find the efficiency of exosomes as a delivery vehicle, the expression level of some miR-145 target genes, including Rho-Associated Coiled-Coil Containing Protein Kinase 1 (ROCK1), Erb-B2 Receptor Tyrosine Kinase 2 (ERBB2), Matrix Metalloproteinase 9 (MMP9), and Tumor Protein p53 (TP53), was compared in all treatment groups (T-47D cells treated by miR-145-transfected MSCs and their derivatives or their backbone) and control group (untransfected T-47D cells) using real-time PCR. RESULTS: The obtained data represented the inhibitory effect of miR-145 on apoptosis induction and metastasis in both direct miR-treated groups. However, exosome-mediated delivery caused an improved anticancer property of miR-145. CONCLUSION: Restoration of miR-145 using MSC-Exo can be considered a potential novel therapeutic strategy in breast cancer in the future.

Inhibitory effects of probiotic Bacillus coagulans against MCF7 breast cancer cells.

BACKGROUND AND OBJECTIVES: Secondary metabolites in the supernatants of probiotic microorganisms have shown anticancer effects. The present study was aimed to investigate the cytotoxicity of Bacillus coagulans supernatants and their role in apoptosis induction in MCF7 cancer cells. MATERIALS AND METHODS: The inhibition of MCF7 cancer cells by Bacillus coagulans supernatants was assessed by MTT assay at three exposure times of 24, 48, and 72 h. Apoptosis induction was explored by flow cytometry while the expression levels of bax, caspase 3, caspase 9, and bcl2 were examined by real-time PCR and compared with normal HFF cells. RESULTS: Bacillus coagulans supernatants exhibited inhibitory effects on MCF7 cells in a concentration-dependent and time-dependent manner; while lower cytotoxic effects were observed in normal HFF cells. The increase in the expression of bax, caspase 3, and caspase 9 genes and the decrease in the anti-apoptotic gene of bcl2, along with the flow cytometry results, confirmed the induction of apoptosis in the cancer cells. CONCLUSION: Regarding the cytotoxic influence of Bacillus coagulans supernatants against breast cancer cells, this bacterium can be considered as a potential candidate for a novel therapeutic strategy with lower side effects which of course requires further investigations.

Genital infections and reproductive complications associated with Trichomonas vaginalis, Neisseria gonorrhoeae, and Streptococcus agalactiae in women of Qom, central Iran.

BACKGROUND: Trichomonas vaginalis (T.vaginalis) and Neisseria gonorrhoeae (N.gonorrhoeae) are two most common non-viral sexually transmitted infections in the world. No data are available regarding the epidemiology of genital infections in women of Qom, central Iran. OBJECTIVE: Epidemiological investigation of sexually transmitted infections in genital specimens of women referred to the referral gynecology hospital in Qom, central Iran. MATERIALS AND METHODS: Genital swab specimens were collected from women volunteers and used for identification of bacterial and protozoal infections by conventional microbial diagnostics, porA pseudo gene LightCycler® real-time PCR (for N.gonorrhoeae) and ITS-PCR (for T.vaginalis). RESULTS: Of 420 volunteers, 277 (65.9%) had genital signs/symptoms, including 38.3% malodorous discharge, 37.9% dyspareunia, and 54.8% abdominal pain. Totally, 2 isolates of Streptococcus agalactiae were identified. Five specimens (1.2%) in Thayer-Martin culture and 17 (4.1%) in real-time PCR were identified as N.gonorrhoeae. Fifty-four specimens (12.9%) in wet mount, 64 (15.2%) in Dorset's culture, and 81 (19.3%) in ITS-PCR showed positive results for T.vaginalis. Five mixed infections of T.vaginalis+ N.gonorrhoeae were found. The risk of T.vaginalis infection was increased in women with low-birth-weight (p=0.00; OR=43.29), history of abortion (p=0.00; OR=91.84), and premature rupture of membranes (PROM) (p=0.00; OR=21.75). The probability of finding nuclear leukocytes (p=0.00; OR=43.34) in vaginal smear was higher in T.vaginalis infection. CONCLUSION: The significant prevalence of trichomoniasis and gonorrhea emphasizes the need for accurate diagnosis and effective surveillance to prevent serious reproductive complications in women.

Analysis of killer cell immunoglobulin-like receptors (KIRs) and their HLA ligand genes polymorphisms in Iranian patients with systemic sclerosis.

Genetic factors have a great role in the pathogenesis of autoimmune diseases by cooperating with environmental stimuli. Killer immunoglobulin-like receptors (KIRs) are cell surface proteins on NK cells whose association with major histocompatibility complex-I regulates their killing function. The aim of this study was to provide information on the possible association between KIR and human leukocyte antigen (HLA) genes with systemic sclerosis disease in Iranian population. A total of 279 systemic sclerosis patients and 451 healthy controls were enrolled in this case-control study in order to determine the presence or absence of 19 KIR genes and 6 specific HLA class I ligands. DNA was analyzed by polymerase chain reaction using the specific sequence primer method (PCR-SSP). Among 11 discovered KIR genotypes, 6 genotypes showed a considerable role and 4 genotypes could preclude the risk of systemic sclerosis (SSc) disease. The gene-gene interactions were also analyzed, and significant confounding effects were seen between involved genes in these two combinations: "KIR3DL1; HLA-BW4-Thr80" and "KIR3DL1 -HLA-BW4-A1." None of single KIR genes showed significant effect on the risk of SSc. We conclude that there is an important relationship between KIR genes and their HLA ligands with incidence rate of systemic sclerosis in Iranian population. The powerful role of a number of discovered KIR/HLA compounds such as activating KIR genotype 3 and HLA-BW4-A1 confirmed the provocative hypothesis of the interplay between activating or inhibitory KIR genes with HLA ligands as a critical index of systemic sclerosis predisposition.

Britannin, a sesquiterpene lactone, inhibits proliferation and induces apoptosis through the mitochondrial signaling pathway in human breast cancer cells.

Induction of apoptosis in cancer cells can be a promising treatment method in cancer therapy. Naturally derived products had drawn growing attention as agent in cancer therapy. The main target of anticancer drugs may be distinct, but eventually, they lead to identical cell death pathway, which is apoptosis. Here, we indicated that britannin, a sesquiterpene lactone isolated from Asteraceae family, has antiproliferative activity on the MCF-7 and MDA-MB-468 human breast cancer cells. Annexin V/propidium iodide (PI) staining, Hoechst 33258 staining, and caspase-3/9 activity assay confirmed that britannin is able to induce apoptosis in MCF-7 and MDA-MB-468 cells. The Western blot analysis showed that the expression of Bcl-2 was noticeably decreased in response to britannin treatment, while the expression of Bax protein was increased, which were positively correlated with elevated expression of p53. Moreover, britannin also increased reactive oxygen species (ROS) generation which in turn triggered the loss of mitochondrial transmembrane potential (ΔΨm) and the subsequent release of cytochrome c from mitochondria into cytosol. Taken together, these results suggest that britannin inhibits growth of MCF-7 and MDA-MB-468 breast cancer cells through the activation of the mitochondrial apoptotic pathway and may potentially serve as an agent for breast cancer therapy.

High carrier frequency of the GJB2 mutation (35delG) in the north of Iran.

OBJECTIVE: Mutations in the GJB2 gene are a major cause of autosomal recessive and sporadic non-syndromic hearing loss in many populations. A single mutation of this gene (35delG) accounts for approximately 70% of mutations in Caucasians with a carrier frequency of 2-4% in Europe. This study aims to determine the rate of 35delG carrier frequency in Iran. METHODS: Genomic DNA was extracted from a total of 550 unaffected unrelated subjects from 4 provinces of Iran following the standard phenol chloroform procedure. The one base pair deletion (35delG) was analysed using a nested PCR procedure; 35delG mutation carriers were subsequently confirmed by sequence analysis. Moreover, using the Binomial probability distribution, we compared the 35delG carrier frequency of Iranian population with the various Middle Eastern and overall European populations. RESULTS: Of the four populations studied, we found a high carrier frequency of 2.8% in Gilan province in the north of Iran. The overall 35delG carrier frequency was found to be 1.25% in the populations studied (our present and previous data) which is similar to the overall 35delG carrier frequency detected in Middle Eastern populations, but Significantly lower than that identified in European populations.