Expression of proliferation-related genes in BM-MSC-treated ALL cells in hypoxia condition is regulated under the influence of epigenetic factors in-vitro.

Mesenchymal stem cells affect ALL cell biology under hypoxic conditions. We studied survival, proliferation, expression, and promoter methylation levels of essential genes involved in expanding MOLT-4 cells co-cultured with BM-MSC under the hypoxic condition. Here, MOLT-4 cells were co-cultured with BMMSCs under hypoxic conditions. First, the apoptosis rate was evaluated by Flow cytometry. Then, MOLT-4 cells' proliferation rate was assessed using MTT assay, and the expressions and methylation rates of genes were determined by qRT-PCR and MS-qPCR, respectively. The results showed that although MOLT-4 cells proliferation and survival rates were reduced under hypoxic conditions, this reduction was not statistically significant. Also, we showed that hypoxic conditions caused upregulation of candidate genes and affected their methylation status. Besides, it was revealed that Pontin was downregulated, while KDM3A, SKP2, and AURKA had an upward trend in the presence of MOLT-4 cells plus BM-MSC. The co-culture of leukemia cells with BMMSCs under hypoxic conditions may be a potential therapeutic approach for ALL.

Up-regulation of KISS1 as a novel target of Let-7i in melanoma serves as a potential suppressor of migration and proliferation in vitro.

Melanoma is a kind of skin cancer that is begun by the alteration of melanocytes. miRNAs are small non-coding RNA molecules that regulate a variety of biological processes. KISS1, the metastasis-suppressor gene, encodes kisspeptins which inhibits migration and proliferation of cancers. This study was aimed to determine the role of Let-7i and KISS1 in melanoma cell migration and proliferation. At first, the expression of Let-7i and KISS1 was determined in patients with melanoma. In the in vitro part of the study, Let-7i mimics were transfected and the impact of its restoration on target gene expression, proliferation, migration and apoptosis of SK-MEL-3 melanoma cell line was assessed by real-time PCR and Western blotting, MTT assay, wound-healing assay and flow cytometry, respectively. Besides, KISS1 inhibitor siRNA alone and along with Let-7i was transfected to determine their probable correlation. The results revealed that either Let-7i or KISS1 were down-regulated in patients with melanoma. The results obtained from the in vitro part of the study revealed that restoration of Let-7i reduced the expression of metastasis- and proliferation-related target genes. Moreover, it was revealed that up-regulation of Let-7i attenuated migration and proliferation capability of SK-MEL-3 cells. Besides, it was demonstrated that Let-7i restoration induced apoptosis in melanoma cells. More importantly, the KISS1 inhibitor caused a prominent cell migration and proliferation, attenuated by Let-7i re-expression. To sum up, the present study revealed the impressive role of Let-7i restoration along with its correlation with KISS1 on melanoma carcinogenicity which may be applicable in future in vivo studies.

miR-193a-5p as a promising therapeutic candidate in colorectal cancer by reducing 5-FU and Oxaliplatin chemoresistance by targeting CXCR4.

Colorectal cancer (CRC) is one of the most common causes of cancer-related deaths worldwide. The role of microRNAs (miRNAs/miRs) as small (19-25 nucleotides in length) non-coding RNA molecules that modify gene expression has been shown in several types of cancer. 5-Fluorouracil (5-FU) and oxaliplatin (Ox) are two common chemotherapeutic agents used to treat cancer. The present study aimed to evaluate the expression levels of miR-193a-5p in CRC, and its effect on the C-X-C Motif Chemokine Receptor 4 (CXCR4) target gene alone and in combination with chemotherapeutic drugs, to determine its possible role in chemoresistance. CRC tissues and adjacent non-cancerous tissue were obtained from 67 patients who had undergone surgery to determine the expression levels of miR-193a-5p and CXCR4. Subsequently, qPCR and Western blotting were performed to determine the effect of miR-193a-5p and chemotherapy drugs on CXCR4. َAlso, MTT assay, and flow cytometry was performed to determine their role in cell viability and apoptosis. Besides, the relationship between miR-193a-5p and CXCR4 with patients' clinical features was investigated. The results of the present study showed that miR-193a-5p was significantly downregulated, whereas CXCR4 was significantly upregulated in tumor tissues obtained from patients with CRC compared with the adjacent non-tumor healthy controls. In addition, the upregulation of miR-193-5p reduced the expression levels of CXCR4, particularly in combination with 5-FU and OX. Besides, using rescue experiments, the present study showed that miR-193a-5p replacement was able to suppress CXCR4-induced CRC cell proliferation by directly targeting CXCR4. Furthermore, there was a significant association between miR-193a-5p and CXCR4 with certain clinicopathological characteristics, particularly with metastasis-related features. These results suggest that miR-193a-5p serves a tumor-suppressive function in CRC and can directly target CXCR4 and decrease its mRNA and protein expression levels. Additionally, miR-193a-5p in combination with 5-FU and Ox potentiated reducing CXR4 expression, which may reveal its contribution to tumor chemoresistance. In conclusion, miR-193-5p may be applicable as a prognostic and diagnostic marker, and also serve as a therapeutic factor by reducing CXCR4 in combination with chemotherapeutic drugs.

Frequency of Hepatitis C Virus Genotypes in Patients Who Had Hepatocellular Carcinoma in Gastroenterological Hospitals of Tabriz.

BACKGROUND: Generally, hepatitis C has been identified as one of the major health issues that about 3% of the world's population have been threatened and affected by it (about 170 million people), and also, it can be considered a factor in acute and chronic hepatitis. METHODS: The aim of this study is to determine the prevalence of HCV genotypes in Azerbaijan patients. In this study, sampling was done on the referred patients to the hospitals (Mahallati and Behbud Hospital). RNA was extracted after isolation of plasma, and then, after the synthesizing of cDNA, the sample was carried out to the laboratory for performing the real-time PCR in order to determine the genotypes. RESULTS: The evaluation of HCV genotypes in positive plasma samples showed that dominant subsets were remarkable and the mean age of the patients was 37/3 ± 11/8 (in the age range of 2-63). Among the 235 patients,139 of them (59%) were male. Statistically, the average number of women was more than men (T test, P < 0/05). 1b genotype was reported 70% in the patients above 40 years old, and also, it was reported as 71/6% in the patients under 40 years old that was not statistically significant. The incidence of serotype 3a was higher among the patients younger than 40 years old (3a was 18.1% vs. 15%), and this serotype was prevalent among men (3a was 18.7% vs. 14.6%), which was statistically significant. CONCLUSION: The findings indicate that among Azerbaijan's patients with chronic hepatitis C, genotypes 1b (71.1%) and 3a (17%) were dominant.

Prognostic and Diagnostic Values of miR-506 and SPON 1 in Colorectal Cancer with Clinicopathological Considerations.

PURPOSE: One of the most common cancers in the world is colorectal cancer, which has increased significantly in recent decades. In the carcinogenicity process in the colon, there are genes involved in various cellular processes, such as cell cycle, apoptosis, and cell migration. According to studies carried out, both miR-506 and SPON 1 genes are involved in the process which initiates and promotes cancer. In this study, alterations in the expression of target genes from the viewpoint of carcinogenicity were studied and evaluated. METHODS: Fifty tumor tissues and normal marginal tissue were collected from patients who were undergoing colorectal cancer surgery. After the extraction of RNA, the real-time PCR method was performed to evaluate the gene expression. Also, alterations of gene expression in response to defined amounts of chemotherapeutic drugs (IC50) were evaluated. P < 0.05 was considered statistically significant. RESULTS: The relative expression level of miR-506 in tumor tissues was significantly decreased in comparison with healthy marginal tissues (P = 0.044). On the other hand, the SPON1 gene expression level was decreased too in tumor tissues in comparison with healthy marginal tissues (P = 0.019). There was also a significant relationship between the expression of target genes and clinicopathological involvement. However, there was no significant alteration in these genes along with the chemotherapeutic drug. CONCLUSION: These findings suggest that the relative expression of miR-506 and SPON 1 gene can be considered as a diagnostic or predictive biomarker for colorectal cancer. However, further studies on protein levels should be conducted.

Heat Shock Proteins Regulating Toll-like Receptors and the Immune System could be a Novel Therapeutic Target for Melanoma.

Melanoma is a serious type of skin cancer, which develops in melanocyte cells. Although it is less common than some other skin cancers, it can be far more dangerous if not treated at an early stage because of its ability to spread rapidly to other organs. Heat shock proteins (HSP) are intracellular molecular chaperones of naive proteins, which are induced in response to stressful conditions. HSP is released into the extracellular milieu and binds to Toll-like receptors (TLRs) to regulate immune responses, such as cytokine and chemokine release. HSPs can release and bind to tumor-specific antigens, with cross-presentation of major histocompatibility complex (MHC) class I antigens. TLRs are innate immune system receptors, involved in the melanoma growth pathway through HSP activation. Melanocytes express TLR4 and TLR9 to modulate immune responses. Many TLR ligands are considered as proper adjuvant candidates, as they can activate dendritic cells. Targeting some TLRs, such as TLR7 and TLR9, is an available option for treating melanoma. In this review, we aimed to determine the relationship between TLRs and HSP groups in melanoma.