Functional evaluation of EZH2 Gene Silencing Effect on β-catenin Marker Expression in YM-1 Squamous Cell Carcinoma Cell Line of Esophageal Cancer

Subject Areas : Journal of Animal Biology

Fatemeh Nourmohammadi ¹ , Mohammad Mahdi Forghani Fard ² , Vajihe Zarrinpour ³ , Mohammad Reza Abbaszadegan ⁴

1 - Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran
2 - Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran
3 - Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran
4 - Medical Genetics Research Center, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Received: 2023-07-23 Accepted : 2023-07-31 Published : 2024-02-20

Keywords: β-catenin, EZH2, shRNA, Esophageal squamous cell carcinoma,

Abstract :

Esophageal cancer is the sixth cause of cancer death in the world and the eighth most common cancer in the world. One of the most characteristic features of this cancer is the resistance to chemo-radiotherapy and the high percentage of recurrence in patients. Beta-catenin marker is one of the main factors of metastasis and invasion in patients. EZH2 gene is a regulatory factor of gene expression and increases the proliferation of tumor cells and maintains the pluripotency of stem cells. EZH2 is aberrantly expressed in a variety of malignant cancers. β-Catenin is a structural component of adherens junctions along with cadherin and the main signaling agent of the Wnt pathway in the nucleus. Aberrant expression of beta-catenin often leads to cancer and metastasis. In this study, the EZH2 gene silencing vector was cloned in Escherichia coli bacteria and then extracted, and the EZH2 gene was silenced in YM-1 esophageal cancer cells. Then RNA extraction and cDNA synthesis were evaluated after confirming EZH2 gene silencing, beta-catenin expression using Real-time PCR technique. Cell migration test was used to check the invasion rate of cells after genetic manipulation. The results showed a decrease in beta-catenin expression in cell lines induced by EZH2 silencing. EZH2 silencing significantly reduced the growth and migration rate of esophageal cancer cells. The results indicate the role of EZH2 gene in regulating beta-catenin gene expression in esophageal squamous cell cancer. Considering that tumor spread is one of the most important factors in the malignancy of ESCC cancer, and on the other hand, these features of esophageal cancer cause premature death of patients due to metastasis and invasion, finding an effective marker in controlling cancer stem cells can be promising for the treatment of ESCC patients.

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