The Prevalence of P53 Mutations in Laryngeal Cancer in Kerman

Abstract: Background &Aims: Laryngeal cancer is the second common cancer of respiratory tract, following the lung cancer. Carcinogenesis is a complex multistage process; molecular genetics has provided the evidence that activation of proto-oncogene and loss or inactivation of tumor suppressor genes (TSG) are involved in a large number of malignancies. One of the earliest significant tumor suppressor genes identified in head and neck squamous cell carcinoma (HNSCC) was P53 have a role in growth suppression activities. Thus, when P53 is deleted or silenced, the cell develops a selective growth advantage and becomes a cancer. Mutations in P53 are correlated with poorer survival and response to treatment. The aim of this study was to survey the prevalence of P53 gene mutation in patients with laryngeal cancer and to select an appropriate method of treatment. Methods: The samples were 52 patients with laryngeal cancer diagnosis have been treated by surgery. Investigation of TP53 mutation where performed by multiple ligation probe amplification (MLPA) technique which analyze the full length of gene from exon 1 to 12. Results: The TP53 mutation was discovered in 80.8 percent of samples. By contrast between two main forms of mutation (i.e. deletions and duplications), we found that the deletions mostly occurred within exons 1, 3, 6, 9 and 12 by 59.6 percent and duplications observed in exons 1, 2, 7, 8 and 11 by 21.2 percent. Conclusion: Considering our results and reminding this fact that nowadays the definitive diagnosis of laryngeal cancer is made using biopsy and pathology techniques, we suggest that all biopsy specimens should be tested and those confirmed positive for TP53 mutations need some further decisions by physicians. Keywords: Mutation, P53, Cancer, Multiple ligation probe amplification (MLPA), Laryn

Prevalence of Nucleotide Alterations of EGFR Gene in Patients with Esophageal Squamous Cell Carcinoma in Kerman

Abstract: Background & Aims: Esophageal Cancer is the sixth fatal cancer in the world. Squamous and adenocarcinoma account for 95% of esophageal cancer. The expression of EGFR has a role in the pathophysiology of epidermal-based malignancies such as esophageal cancer. EGFR is also an important criterion in the evaluation of disease staging and prognosis. The aim of this study was to survey the prevalence of EGFR gene mutations in patients with esophageal cancer by MLPA Technique. Methods: A total of 60 parafinated blocks from patients with esophageal cancer were investigated for the presence of EGFR mutations by MLPA technique. Results: EGFR mutation was discovered in 82 percent of samples of which 52% were deletion mostly seen in exon 2 (52%) and duplication mostly seen in exon 27 (54%). In some cases deletion and/or duplication were seen in more than one exon simultaneously. Conclusion: With regard to the obtained results and since the definitive diagnosis of esophageal cancer is made by biopsy and pathology techniques, it is suggested that all biopsy specimens be detected for EGFR mutations, particularly on exons 2 and 27 in order to achieve a noninvasive molecular diagnostic technique in future. Keywords: Esophageal neoplasm, EGFR, Mutation, MLP

Transplantation of differentiated umbilical cord mesenchymal cells under kidney capsule for control of type I diabetes in rat

Nowadays, stem cells have been introduced as an appropriate source of regenerative medicine for treatment of type I diabetes. Human umbilical cord matrix-derived mesenchymal cells (hUCMC) have successfully been differentiated into insulin producing cells. The isolated hUCM cells were characterized by the expression of stem cell surface markers and by differentiation into adipocytes and osteocytes. The hUCMCs were cultured with different concentrations of neural conditional medium (NCM) and were induced to differentiate into insulin producing cells (IPCs). As 60% NCM concentration resulted in higher nestin and PDX1 expression, the cells were first exposed to 60% NCM and were then induced for IPCs differentiation. PDX1 and insulin gene expression was evaluated in the treated cells. Also, the secretion capacity of the IPCs was assessed by glucose challenge test. IPCs were transferred under the rat kidney capsule. Blood glucose level, weight gain and immunohistochemistry assessments were done in the treated animals. hUCMC expressed mesenchymal cell surface markers and successfully differentiated into adipocytes and osteocytes. Higher NCM concentration resulted in higher PDX1 and nestin expression. The IPCs expressed insulin and PDX1. IPCs were detectable under the kidney capsule 2 months after injection. IPCs transplantation resulted in a sharp decline of blood sugar level and less weight loss. Differentiated hUCM cells could alleviate the insulin deprivation in the rat model of type I diabetes. In addition, higher NCM concentration leads to more differentiation into IPCs and more nestin and PDX1 expression. Kidney capsule can serve as a suitable nominee for IPCs transplantation

Deletion/duplication mutation screening of TP53 gene in patients with transitional cell carcinoma of urinary bladder using multiplex ligation-dependent probe amplification

Bladder cancer is a molecular disease driven by the accumulation of genetic, epigenetic, and environmental factors. The aim of this study was to detect the deletions/duplication mutations in TP53 gene exons using multiplex ligation-dependent probe amplification (MLPA) method in the patients with transitional cell carcinoma (TCC). The achieved formalin-fixed paraffin-embedded tissues from 60 patients with TCC of bladder were screened for exonal deletions or duplications of every 12 TP53 gene exons using MLPA. The pathological sections were examined by three pathologists and categorized according to the WHO scoring guideline as 18 (30%) grade I, 22 (37%) grade II, 13 (22%) grade III, and 7 (11%) grade IV cases of TCC. None mutation changes of TP53 gene were detected in 24 (40%) of the patients. Furthermore, mutation changes including, 15 (25%) deletion, 17 (28%) duplication, and 4 (7%) both deletion and duplication cases were observed among 60 samples. From 12 exons of TP53 gene, exon 1 was more subjected to exonal deletion. Deletion of exon 1 of TP53 gene has occurred in 11 (35.4%) patients with TCC. In general, most mutations of TP53, either deletion or duplication, were found in exon 1, which was statistically significant. In addition, no relation between the TCC tumor grade and any type of mutation were observed in this research. MLPA is a simple and efficient method to analyze genomic deletions and duplications of all 12 exons of TP53 gene. The finding of this report that most of the mutations of TP53 occur in exon 1 is in contrast to that of the other reports suggesting that exons 5-8 are the most (frequently) mutated exons of TP53 gene. The mutations of exon 1 of TP53 gene may play an important role in the tumorogenesis of TCC