Evaluation of apoptosis induced by CTLA-4 inhibition in Esophageal cancer cell

Abstract

Esophageal cancer is a type of cancer that develops in the esophagus or trachea (a part of the digestive tract). The esophagus is responsible for transporting food to the stomach and is located between the pharynx and the stomach. Symptoms of this disease include difficulty in swallowing and weight loss. CTLA-4 are inhibitory molecules that regulate the immune system, which, by being expressed on the surface of cancer cells, play a role in reducing the function of immune cells. In this study, the aim is to suppress the expression of CTLA-4 genes by using specific siRNA and evaluate the effect of this simultaneous suppression on the growth, migration and apoptosis of cancer cells.

Materials and methods:

First, the cell line (TE-8) (Esophageal cancer) was cultured. Then CTLA4 specific siRNA was transfected into the desired cell line. MTT test was performed to check the cytotoxic effect of transfected siRNA. In order to investigate this gene suppression on the rate of growth and proliferation of cancer cells, genes involved in apoptosis, and DNA degradation (BAX, BCL-2, CAS3, CAS9) were investigated by PCR-RT method. After collecting data in Excell software, data organization was done and then data distribution was checked using Prism software and for normal distribution by Test-T test and for data with non-normal distribution by Whitney test. -Mann was used.

Results: We observed a significant increase in CTLA-4 expression in the TE-8 cell line. Compared to the control group, our results showed a significantly decreased frequency of genes involved in apoptosis such as BCL-2 after adding CTLA-4 siRNA. while genes involved in DNA degradation and apoptosis such as CAS3, CAS9, and BAX showed a significant increase. Also, the data from MTT showed that after adding CTLA-4 siRNA, the rate of growth and proliferation of cancer cells decreased.