ABSTRACT
Background: Human leukocyte antigen G belongs to the family of non-classical HLA class I genes, its expression considered an important immune escape mechanism of cancer cells. The polymorphisms in the 3'-untranslated region (UTR) region of HLA-G influence the magnitude of the protein by modulating HLA-G mRNA stability. We hypothesised links between any of eight (UTR) single nucleotide polymorphisms (SNPs) and their haplotype of the HLA-G gene with breast cancer. Materials and Methods: Peripheral blood DNA from 100 patients affected by breast cancer and 100 controls was PCR sequenced for genotyping of 25 HLA-G 3'-UTR regions, including rs371194629 (+2960), rs1707 (+3003), rs1710 (+3010), rs17179101 (+3027), rs1063320 (+3142), rs9380142 (+3187), rs1610696 (+3196), and rs1233331 (+3227). Results: The 14-bp deletion (p = 0.01), and the +3010 (p = 0.021), +3142 (p = 0.006) and +3187 (p = 0.046) variants were significantly more prevalent in patients than in controls. In combining these data, two haplotypes of all eight SNPs and deletion/insertion (UTR-1 and UTR-4) are associated with breast cancer. Conclusion: Certain variants in the 3-UTR, and their combination as a haplotype, of the HLA-G gene are linked to breast cancer.
Subject(s)
Breast Neoplasms , Genes, MHC Class I , 3' Untranslated Regions/genetics , Breast Neoplasms/genetics , Female , Gene Frequency , Genetic Predisposition to Disease , HLA-G Antigens/genetics , Haplotypes , Humans , Polymorphism, Single Nucleotide/geneticsABSTRACT
BACKGROUND: Altered expression of several long non-coding RNAs (lncRNAs) has been described in numerous malignancies, including breast cancer, and some may have a role in carcinogenesis. We hypothesised differences in the expression of lncRNA MCM3AP-AS1 in breast cancer tissues compared to nearby healthy tissues and potential links with clinical features. METHODS: We tested our hypothesis in 102 pairs of breast cancer tumours and adjacent non-tumour tissues from female patients. After RNA extraction, cDNA synthesis was performed for all specimens. The differential gene expression was assessed using Quantitative Real-Time PCR Technique. RESULTS: There was a significant overexpression of the lncRNAs in tumour tissues as compared with their adjacent non-tumour tissues (P < 0.001). Expression was significantly linked with the tumour oestrogen receptor expression (P = 0.023) and tumour progesterone receptor expression (P < 0.001). ROC analysis showed an AUC of 0.67 (95% CI 0.60-0.75) (P < 0.001) with sensitivity and specificity of 58% and 76%, respectively. CONCLUSION: The lncRNA MCM3AP-AS1 may be a novel breast cancer lncRNA with high expression levels in breast cancer patients' tissue. Further investigations are needed to confirm its uses as a potential molecular marker and therapeutic target.
Subject(s)
Breast Neoplasms , MicroRNAs , RNA, Long Noncoding , Acetyltransferases , Breast Neoplasms/genetics , Female , Humans , Intracellular Signaling Peptides and Proteins , RNA, Long Noncoding/genetics , ROC CurveABSTRACT
Calprotectin is a heterodimeric EF-hand Ca2+ binding protein that is typically released by infiltrating polymorphonuclear leukocytes and macrophages. This protein is a key player linking inflammation and cancer. Due to the increased levels of calprotectin in different inflammatory diseases and cancer, it is considered as a marker for diagnostic purposes. In this study, we evaluated the mechanism of cell viability and apoptotic-inducing effects of recombinant human calprotectin (rhS100A8/S100A9) on the gastric adenocarcinoma (AGS), the most common type of gastric cancer cell line. AGS cells were exposed to the different concentrations (5-100 µg/ml) of calprotectin for 24, 48, and 72 h, and cell viability was assessed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptotic-inducing effects of calprotectin were evaluated by sub-G1 cell cycle assay and Annexin V/propidium iodide double staining. Furthermore, real-time polymerase chain reaction and Western blot analysis were performed to evaluate the mechanism of action of calprotectin. Our findings indicated that calprotectin inhibits growth and viability of AGS cells in a time- and dose-dependent manner. The half-maximal inhibitory concentration values were measured as 85.77, 79.14, and 65.39 µg/ml for 24, 48, and 72 h, respectively. Additionally, we found that calprotectin downregulated the expression of antiapoptotic protein Bcl-2 and upregulated proapoptotic protein Bax in a time- and concentration-dependent fashion. Calprotectin also slightly upregulated the expression of extracellular signal-regulated protein kinase 2 (ERK2), while it significantly decreased the levels of phospho-ERK in a time-dependent manner. Overall, these findings indicated that calprotectin has cytotoxicity and apoptosis-inducing effects on AGS cell lines in high concentration by modulating Bax/Bcl-2 expression ratio accompanied by inhibition of ERK activation.
Subject(s)
Adenocarcinoma/genetics , Calgranulin A/genetics , Calgranulin B/genetics , Mitogen-Activated Protein Kinase 1/genetics , Proto-Oncogene Proteins c-bcl-2/genetics , Stomach Neoplasms/genetics , Apoptosis , Cell Line, Tumor , Cell Survival , Down-Regulation/drug effects , Escherichia coli/genetics , Humans , Recombinant Fusion Proteins/geneticsABSTRACT
The objective of this study was to examine the prophylactic and therapeutic effect of whey-cultured Lactobacillus casei (L. casei) in a murine model of colitis. Colitis was induced by intracolonic administration of a mixture of 2,4,6-trinitrobenzenesulphonic acid (TNBS)/absolute ethanol in male Wistar rats. Animals were divided into 5 groups including sham (normal group), control (vehicle-treated), positive control (dexamethasone 1 mg/kg/day, orally), prevention (10(8) cfu L. casei/day, orally, 14 days before induction of colitis), and treatment (10(8) cfu L. casei/day, orally, 14 days after induction of colitis). After 14-days treatment, the animals were sacrificed on the day 15. Distal colons were removed for examining histological and biochemical assays. Biomarkers including TNF-α, myeloperoxidase (MPO), and lipid peroxidation (LPO) were measured in the homogenate of colon. Results indicated an apparent improvement in colon histopathology scores, TNF-α, MPO, and LPO in the treatment group, whereas prevention group did not demonstrate positive efficacy in prevention of colonic damage. It is concluded that L. casei grown in whey culture is very effective in ameliorating both biochemical and histopathological markers of colitis if used post induction of colitis but not if used before induction of colitis. The difference between effects of L. casei when used pre-colitis and post-colitis confirms its mechanism of action as an anti toxic stress agent. Further studies should be made in IBD patients.
