Role of SIZN1 in Esophageal Squamous Cell Carcinoma

Background: Bone morphogenetic proteins are a family of cytokines and growth factors that are involved in tumorigenesis. ZCCHC12 (SIZN1), as a transcriptional coactivator of bone morphogenetic protein signaling, is identified as a positive regulator of central nervous system development during embryogenesis. It positively regulates the CREB and AP1 transcription factors that cooperate with the bone morphogenetic protein signaling pathway. In the present study, SIZN1 mRNA expression was assessed in esophageal squamous cell carcinoma patients. Methods: The levels of SIZN1 mRNA expression in tumor tissues from 50 patients with esophageal squamous cell carcinoma were compared with their corresponding normal margins by using real-time polymerase chain reaction. Results: We observed that 10 out of 50 (20%) cases overexpressed SIZN1, whereas 40 out of 50 (80%) cases showed either normal or under expression of SIZN1. There was a significant correlation between the levels of SIZN1 mRNA expression and tumor depth of invasion (P=0.040). Furthermore, a significant correlation between lymph node metastasis and SIZN1 mRNA expression was observed in esophageal squamous cell carcinoma patients (P=0.036). Conclusion: This study is the first report that has assessed SIZN1 expression in esophageal squamous cell carcinoma patients. SIZN1 can be a potential therapeutic target for primary esophageal squamous cell carcinoma because of its role in the early stages of tumor progression and metastasis

Cancer Stem Cell Markers in Esophageal Cancer

Esophageal carcinoma is one of the most malignant of all tumors, and affected patients have low survival rates. The lack of good prognostic and therapeutic targets indicate the mysterious biology of this cancer. Recently, studies with cancer stem cells (CSCs) revealed some clues for better understanding of cancer biology and development. CSCs are derived from normal stem cells and have essential roles in tumor initiation and development of malignancies, such as esophageal carcinoma. Self-renewal studies in CSCs have improved our understanding of the factors that regulate CSCs behaviour and may result in improved prognostic markers and new therapeutic targets. Abnormal activity of major cell signaling pathways such as Shh, Notch, and Wnt play important roles in converting stem cells from normal to cancerous. This manuscript reviews the importance of several processes in the maintenance of esophageal CSCs and introduces probable useful markers for CSC based ESCC therapy

Telomere shortening associated with increased levels of oxidative stress in sulfur mustard-exposed Iranian veterans

Sulfur Mustard (SM) is the most widely used chemical weapon. It was used in World War 1 and in the more recent Iran-Iraq conflict. Genetic toxicity and DNA alkylation effects of SM in molecular and animal experiments are well documented. In this study, lymphocytic telomere lengths and serum levels of isoprostane F2α were measured using q-PCR and enzyme immunoassay-based methods in 40 Iranian veterans who had been exposed to SM between 1983-88 and 40 non-exposed healthy volunteers. The relative telomere length in SM-exposed individuals was found to be significantly shorter than the non-exposed individuals. In addition, the level of 8-isoprostane F2α was significantly higher in the SM-exposed group compared to controls. Oxidative stress can be caused by defective antioxidant responses following gene mutations or altered activities of antioxidant enzymes. Chronic respiratory diseases and infections may also increaseoxidative stress. The novel finding of this study was a the identification of ‘premature ageing phenotype’. More specifically, telomere shortening which occurs naturally with aging is accelerated in SM-exposed individuals. Oxidative stress, mutations in DNA repair genes and epimutaions may be among the major mechanisms of telomere attrition. These findings may help for a novel therapeutic strategy by telomere elongation or for validation of an exposure biomarker for SM toxicity

Cloning and expression of the V-domain of the CD166 in prokaryotic host cell

Purpose: CD166/ALCAM (Activated leukocyte cell adhesion molecule) as an immunoglobulin is implicated in cell migration. It is also involved in tumorigenesis of CRC (colorectal cancer) and known as a cancer stem cell marker. CD166, as a membrane protein, potentially represents either diagnostic or therapeutic capacities for CRC.Methods: In this study, the sequence of V domain was optimized for expression in prokaryotic host using online tools and cloned into pET-28a plasmid. The recombinant pET28a was transformed into the E. coli BL21DE3 using heat shock method and expression of recombinant V domain was examined using SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis).Results: The results confirmed protein expression of recombinant 22.77 kDa V domains in bacterial expression system.Conclusion: V domain of the CD166 was expressed successfully in E. coli bacteria. This recombinant fragment can be introduced as a suitable diagnostic and therapeutic candidate for screening and cancer-therapy of CRC patients, respectively.

In vitro Cytotoxicity Effect of Lactobacillus casei on Kyse-30 Human Esophageal Cancer Cells

One of the top causes of cancer-related deaths globally is esophageal cancer. In investigations of cell toxicity, the MTT test is one of the most often used cell viability/cytotoxicity assays for cellular metabolic activity. Nowadays, lactobacilli with probiotic effectiveness are now acknowledged as a prophylactic agent against cancer. The anti-tumor product of these bacteria have been designated in numerous studies. This investigation examined the probiotic Lactobacillus casei's in vivo impact on esophageal cancer. The MTT technique was used in this work to evaluate the cytotoxicity of L. casei (supernatant and full cell culture) to 5fu on the cancer cell line Kyse30. L. casei was able to decrease cell survival in supernatant and full cell culture (Kyse30). The possible impact of L. casei, particularly their supernatant, on esophageal cancer was initially evaluated in this research. As a result, lactobacilli species show promise for future research and development as cancer treatments

TWIST1 Plays Role in Expression of Stemness State Markers in ESCC

Background: Stemness markers play critical roles in the maintenance of key properties of embryonic stem cells (ESCs), including the pluripotency, stemness state, and self-renewal capacities, as well as cell fate decision. Some of these features are present in cancer stem cells (CSCs). TWIST1, as a bHLH transcription factor oncogene, is involved in the epithelial–mesenchymal transition (EMT) process in both embryonic and cancer development. Our aim in this study was to investigate the functional correlation between TWIST1 and the involved genes in the process of CSCs self-renewal in human esophageal squamous cell carcinoma (ESCC) line KYSE-30. Methods: TWIST1 overexpression was enforced in the ESCC KYSE-30 cells using retroviral vector containing the specific pruf-IRES-GFP-hTWIST1 sequence. Following RNA extraction and cDNA synthesis, the mRNA expression profile of TWIST1 and the stem cell markers, including BMI1, CRIPTO1, DPPA2, KLF4, SOX2, NANOG, and MSI1, were assessed using relative comparative real-time PCR. Results: Ectopic expression of TWIST1 in KYSE-30 cells resulted in an increased expression of TWIST1 compared to control GFP cells by nearly 9-fold. Transduction of TWIST1-retroviral particles caused a significant enhancement in BMI1, CRIPTO1, DPPA2, KLF4, and SOX2 mRNA expression, approximately 4.5-, 3.2-, 5.5-, 3.5-, and 3.7-folds, respectively, whereas this increased TWIST1 expression caused no change in the mRNA expression of NANOG and MSI1 genes. Conclusions: TWIST1 gene ectopic expression in KYSE-30 cells enhanced the level of cancer stem cell markers’ mRNA expression. These results may emphasize the role of TWIST1 in the self-renewal process and may corroborate the involvement of TWIST1 in the stemness state capacity of ESCC cell line KYSE-30, as well as its potential as a therapeutic target

Correlation of Wnt and NOTCH pathways in esophageal squamous cell carcinoma

There is an inevitable association between cell signaling pathways and tumorigenesis. Wnt and notch pathways play important roles during development and self-renewal. Beside the independent role of such pathways on tumor progression, different cross talks between these pathways through tumorigenesis are emphasized. In this study, we analyzed cross talk between Wnt and NOTCH signaling pathways through assessment of probable correlation between MAML1 and PYGO2 as the main transcription factors of these pathways, respectively in esophageal squamous cell carcinoma (ESCC) patients. Levels of MAML1 and PYGO2 mRNA expression in 48 ESCC patients were compared to the correlated margin normal tissues using real-time polymerase chain reaction (PCR). Eleven out of 48 patients (22.9 %) have shown the concomitant MAML1/PYGO2 over expression in significant correlation with tumor size (p = 0.046) and depth of tumor invasion (p = 0.050). We showed that there is a significant correlation and feedback between these markers during the ESCC progression and metastasis

Inhibitory effect of zinc oxide nanoparticles on pseudomonas aeruginosa biofilm formation

Objective(s): Bacterial biofilm formation causes many persistent and chronic infections. The matrix protects biofilm bacteria from exposure to innate immune defenses and antibiotic treatments. The purpose of this study was to evaluate the biofilm formation of clinical isolates of Pseudomonas aeruginosa and the activity of zinc oxide nanoparticles (ZnO NPs) on biofilm. Materials and Methods: After collecting bacteria from clinical samples of hospitalized patients, the ability of organisms were evaluated to create biofilm by tissue culture plate (TCP) assay. ZnO NPs were synthesized by sol gel method and the efficacy of different concentrations (50- 350 µg/ml) of ZnO NPs was assessed on biofilm formation and also elimination of pre-formed biofilm by using TCP method. Results:The average diameter of synthesized ZnO NPs was 20 nm. The minimum inhibitory concentration of nanoparticles was 150- 158 μg/ml and the minimum bactericidal concentration was higher (325 µg/ml). All 15 clinical isolates of P. aeruginosa were able to produce biofilm. Treating the organisms with nanoparticles at concentrations of 350 μg/ml resulted in more than 94% inhibition in OD reduction%. Molecular analysis showed that the presence of mRNA of pslA gene after treating bacteria with ZnO NPs for 30 minutes. Conclusion: The results showed that ZnO NPs can inhibit the establishment of P. aeruginosa biofilms and have less effective in removing pre-formed biofilm. However the tested nanoparticles exhibited anti-biofilm effect, but mRNA of pslA gene could be still detected in the medium by RT-PCR technique after 30 minutes treatment with ZnO

Ectopic expression of TWIST1 upregulates the stemness marker OCT4 in the esophageal squamous cell carcinoma cell line KYSE30

Abstract Background The transcription factor TWIST1 plays an important role in the epithelial–mesenchymal transition (EMT) process and in the migration, invasion and metastasis of cancer cells. OCT4, which is a homeobox transcription factor, has an important role in the self-renewal potential of cancer cells. Our aim here is to elucidate impact of ectopic expression of TWIST1 on OCT4 gene expression in esophageal squamous cell carcinoma (ESCC). Methods The ESCC line was KYSE30. GP293T cells were transfected with purf-IRES-GFP and pGP plasmids to produce recombinant viral particles. A semi-confluent KYSE30 culture was transduced with the prepared retroviral particles. mRNA extraction and cDNA synthesis were performed from normal KYSE30 cells and those ectopically expressing TWIST1. Expressional analysis of TWIST1 and OCT4 were performed with relative comparative real-time PCR. Results Ectopic expression of TWIST1 in KYSE30 cells was related to its significant overexpression: nearly nine-fold higher in GFP-hTWIST1 KYSE-30 cells than in control GFP cells. This induced expression of TWIST1 caused significant upregulation of OCT4 in GFP-hTWIST1 KYSE-30 cells: nearly eight-fold higher. In silico analysis predicted the correlation of TWIST1 and OCT4 through ETS2. Conclusions Overexpressed TWIST1 can be correlated with upregulation of the cancer stem cell marker OCT4 and the protein may play critical regulatory role in OCT4 gene expression. Since OCT4 is involved in the self-renewal process, the results may suggest a new linkage between TWIST1 and OCT4 in the cell biology of ESCC, highlighting the probable role of TWIST1 in inducing self-renewal