The effect of CTLA4 inhibition on Bladder cancer cells survival

Abstract

Bladder cancer is one of the types of squamous cell cancers. Bladder cancer is the second most common genital cancer. The average age of bladder cancer diagnosis is 65 years. Bladder cancer is more common in men. Bladder cancer is more common in whites than blacks. In Iran, men get bladder cancer four times more than women. 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 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 growth, proliferation and metastasis (MMP13, MMP9, MMP2, BCL-2, BAX) 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 cell line. Compared to the control group, our results showed a significantly decreased frequency of genes involved in metastasis and cell migration such as MMP9, MMP13, MMP2, and BCL-2 after adding CTLA-4 siRNA. while genes involved in apoptosis such as 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.