Differential expression of hsa_circ_0064357 and hsa_circ_0064358 between oral squamous cell carcinoma and oral lichen planus

Background/aims: Reliable biomarkers with high specificity and sensitivity and the potential to discriminate precancerous or early lesions from oral cancer improve scientific assessment and early detection. Dysregulated circRNAs play a critical role in the occurrence and progression of malignant biological behaviors of OSCC. The study of potential diagnostic roles of hsa_circ_0064357 and hsa_circ_0064358 in early diagnostic of precancerous lesions such as OLP to OSCC as the most common type of head-and-neck squamous cell carcinoma (HNSCC) was the focus of present research. Methods: The differential expression of hsa_circ_0064357, hsa_circ_0064358, and RAF1 target gene predicted using CircInteractome and Circbase databases between OSCC (n=30), OLP (n=10) tissues and their adjacent normal tissues were evaluated by qRT-PCR. The potential diagnostic value of circRNAs was identified by receiver operating characteristic (ROC) curve analysis. Results: hsa_circ_0064357 and hsa_circ_0064358 were identified to be lowly expressed, while RAF1 was upregulated in OSCC and OLP tissues more than adjacent normal tissues. Low expression of circRNAs was markedly correlated with TNM stages of OSCC patients. ROC analysis revealed AUC of 0.962 and 0.965 for hsa_circ_0064357 and hsa_circ_0064358, respectively, suggesting that circRNAs can serve as novel diagnostic biomarkers for early detection of OSCC. Conclusion: hsa_circ_0064357 and hsa_circ_0064358 might be involved in the progression and metastasis of OSCC and could be used as promising novel biomarkers for early diagnosis and the clinical monitoring of the malignant transformation of OLP into OSCC

Molecular focus in p63 and correlated human diseases

The p63 gene, a member of the p53 gene family, is expressed into at least six protein isoforms.The transcription factor p63 is a homologue of the tumor suppressor p53. Unlike p53, which is dispensable for normal development, p63 is critical for the development of stratified epithelial tissues such as epidermis, breast, and prostate. p63, , is transcribed from two different promoters giving rise to two different proteins: p63, a member of the p53 family, is transcribed from two different promoters giving rise to two different proteins: TAp63 that contains the N-terminal transactivation domain and ΔN that lacks this domain. p63 encodes multiple protein isoforms with both transactivating and transcriptional repressor activities that can regulate a wide spectrum of target genes. p63 is also implicated in tumor formation and progression in stratified epithelia, with evidence for both tumor suppressive and oncogenic properties.

Cloning of Oct3/4 gene in embryonic stem cells

      Embryonic stem cells (ESCs) are pluripotent, self-renewing cells. These cells can be used in applications such as cell therapy, drug discovery, disease modelling, and the study of cellular differentiation. In this experimental study embryonic stem cells cultured in the laboratory and were amplified. Total RNA was extracted from cells and converted to cDNA. The replication factor Oct3/4 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into the pTZ57R/T vector. Legated product had been transformed into susceptible bacteria and transformed bacteria were screened on a selective medium. Plasmids extracted from bacteria and enzyme digestion to confirm the sequencing was performed. Results of enzyme digestion were sequenced. Cloned gene can prepare a gene cassette to produce stem cells from somatic cell

Evaluation of Changes in miR-7113-3p, miR-6721-5p and MAP2K1 gene expressions in tumor and normal tissues of patients with oral cancer

Backgrounds: Squamous cell carcinoma of the oral cavity (OSCC) is one of the most common cancers in the Head and Neck Squamous Cell Cancer (HNSCC) group. The increasing frequency of oral carcinomas and their late-stage appearance is a major worldwide health concern. MicroRNAs (miRNAs) play an important role in cancer growth and progression, as the available relevant data indicate. However, no information is available about the part miR-7113-3p and miR-6721-5p takes in OSCC. In the present study, the expression of MAP2K1, miR-7113-3p, and miR-6721-5p was examined to determine their possible biological role in the advancement of oral squamous cell carcinoma. Methods: Quantitative Real-Time PCR was applied to investigate the mRNA expression of MAP2K1, miR-7113-3p, and miR-6721-5p in fresh frozen OSCC tissues and adjacent normal fresh frozen tissues of 30 patients and then, the relationship between MAP2K1 Expression and clinical parameters was studied. Results: MAP2K1 expression dramatically increased in tumor tissues compared to the normal tissues, while miR7113-3p and miR-6721-5p expression significantly decreased. Furthermore, a statistical correlation of p=0.04 was also observed between increased MAP2K1 expression and Perineural invasion. In addition, the downregulation of miR-7113-3p was positively correlated with overexpression of MAP2K1 (p=0.0218) and a negative correlation was observed between downregulation of miR-6721-5p and overexpression of MAP2K1 (p=0.7771). Conclusion: Based on the findings, miR-7113-3p and miR-6721-5p might be prospective biomarkers for OSCC patients, and can be utilized to detect OSCC at an early stage of its diagnosis. MAP2K1 overexpression is linked to the development of OSCC and Perineural invasion

Effect of Dioscorea extract on Bax and Bcl-2 gene expression in MCF-7 and HFF cell lines

Abstract Background In cancer cells, the balance between proliferation and apoptosis is disturbed. There is a direct relationship between gene expression and the process of apoptosis. The two genes involved in apoptosis are Bax and Bcl-2, and it is now well established that some plant compounds can alter the expression of genes. The aim of this study is to determine the rate of change in the expression of these genes in the cell line MCF-7 treated with Dioscorea extract for 24, 48 and 72 h. For this purpose, the plant extract was prepared by Soxhlet method and diluted in different concentrations. MCF-7 and HFF cell lines were treated in three replicates with different concentrations of the extract at intervals of 24, 48, and 72 h. To evaluate the toxicity of the extract, the MTT assay was performed and the IC50 value was calculated. Both cell types were cultured at IC50 concentration with three treatments and three replicates. RNA extraction, cDNA synthesis and real-time PCR were then performed. Flow cytometry was performed to further confirm apoptosis. Results MTT results showed that 72 h treatment with Dioscorea extract in IC50 concentration had the greatest effect on the death of MCF-7 cancer cells, while the cells of the control cell line remained healthy. The results of the study of gene expression changes showed that when treated with the plant extract for 24 h, the increase in Bax gene expression and the decrease in Bcl-2 gene expression were not statistically significant. At 48-h treatment, the decrease in Bcl-2 expression was not statistically significant, whereas the increase in Bax expression, which was 2.1 times, was statistically significant. When treated with the plant extract for 72 h, Bax expression increased 2.72 times and Bcl-2 gene expression decreased 0.67 times. Flow cytometry showed that 72-h treatment with plant extract at a concentration of 438.35 µg/ml was the most effective treatment for MCF-7 cancer cell death. Conclusions The expression ratio of Bax gene to Bcl-2 is equal to 4.06, which indicates the induction of more apoptosis by treatment with plant extract