جستجوی مقالات مرتبط با کلیدواژه « Touchdown PCR » در نشریات گروه « پزشکی »

Brain tumors are considered to be one of the most dangerous types of cancer, despite decades of research and treatment efforts. The activation of the telomerase enzyme is a critical factor in the immortality of these cells. Mutations in the promoter region of the TERT gene, particularly in the promoter hot spots, can influence the binding of activating transcription factors and inhibit the binding of negative regulatory factors of the TERT gene, ultimately regulating the gene's expression. This study aimed to identify and investigate the mutations of the TERT gene promoter region in glioblastoma, a malignant brain tumor, using bioinformatics.

A study was conducted using the case-control method where 35 patients with glioblastoma multiform brain tumor were examined along with 40 people who were examined as control samples. In this research, the Touchdown PCR technique and direct DNA sequencing were used to detect mutations in the promoter region of the TERT gene in patients with glioblastoma. Furthermore, bioinformatic analyses were conducted to investigate the pathogenic effect of nucleotide changes in this gene region.

In the core region of the TERT gene promoter, three-point mutations were identified (c.*146C>T، c.*144C>G and c.*143G>C). Two of these mutations were novel and have been observed for the first time in glioblastoma multiform. The remaining mutation was located in the promoter core's hot spot of the gene. This mutation was known as c.*146C>T.

In this research, the harmful impact of TERT promoter region mutations as a biomarker for glioblastoma was examined. These findings suggest that mutations in regulatory regions, as well as coding sequences, could be significant contributory factors in the process of carcinogenesis.

Approximately 3% of the human genome is rich in GCs. These regions are often found in the promoter of genes, especially housekeeping genes and tumor suppressor genes. Amplification of these GC-rich regions can be challenging. Because the stability of GC-rich DNA sequences is higher, secondary structures are easily formed in these regions. Type 4 non-medullary thyroid carcinoma has been linked to the FOXE1 gene as a risk factor. FOXE1 belongs to a large family of transcription factors principal for the development of the thyroid gland's morphology.

With a high proportion of GC, a portion of the FOXE1 gene sequence cannot be amplified using the usual polymerase chain reaction. This work is an experimental study that deals with this issue.

The results of the present study showed that the Touchdown polymerase chain reaction, in combination with CO-amplification materials such as betaine and Dimethyl Sulfoxide (DMSO), is effective in amplifying the sequence of this region of the FOXE1 gene by destruction of the secondary structures formed in the sequence and increasing the reaction product.

Using this method, GC-rich regions in additional genes with a similar degree of GC as the FOXE1 gene can be amplified.