Evaluation of the Expression of ID1 Gene in AGS Cells Transfected with pFLAG-CMV3-tagD Recombinant Vector

• Main Authors: Marzieh Rostam zade renani, Abbas Doosti
• Format: Article
• Language: fas
• Published: Ilam University of Medical Sciences 2020-12-01
• Series: Majallah-i Dānishgāh-i ’Ulūm-i Pizishkī-i Īlām
• Subjects: helicobacter pylori; tagd; id1
• Online Access: http://sjimu.medilam.ac.ir/article-1-6275-en.html

Introduction: The ID protein enhances cell proliferation and inhibits differentiation, which determines its association with the process of intestinal tumorigenesis. Helicobacter infection is associated with the expression of different ID genes. This study aimed to investigate the expression of the ID1 gene in AGS cells transfected with the pFLAG-CMV-3-tagD recombinant vector.   Materials & Methods: In the present study, AGS cells were cultured in the RPMI-1640 medium containing 10% bovine fetal serum. Subsequently, these cells were transfected with two pFLAG-CMV-3-tagD or pFLAG-CMV-3 plasmids. Moreover, the plasmid recipient cells were selected by adding 600 mg/LG418. Following that, wwhole-cell RNA was purified using RNX-Plus solution, and the cDNA was synthesized using a kit. The mRNA expression level of ID1 was assessed using q-RT PCR with appropriate primers. Eventually, the expression of each gene was evaluated in SPSS software through an independent t-test. Ethics code: IR. IAU.SHK.REC.1399.026   Findings: The results of RT-PCR confirmed the successful expression of the helicobacter pylori tagD gene in AGS cells. Moreover, the gene expression analysis showed that the ID1 gene expression was significantly decreased in tagD-treated AGS cells, compared to the control cells (P=0.0113).   Discussions & Conclusions: The present study showed that the ID1 gene expression was altered in cells treated with the helicobacter pylori tagD gene. In addition, it seems that the helicobacter pylori tagD gene is involved in this expression change.