Evolution the Effect of linoleic acid on expression of metastatic and angiogenic molecular markers in gastric cancer cell line MKN-45
Abstract
Gastric cancer is a fatal disease characterized by the involvement of gastric tissue. Genetic changes play an important role in the development of gastric cancer. In addition to changes in the level of DNA, changes in the expression level of MicroRNAs are also important in cancer progression. Diet plays an important role in the prevention of this disease and recent studies have confirmed the role of linoleic acid (LA) in some cancers. The aim of the present study was to evaluate the effect of linoleic acid on the expression levels of MMP-2, Talin-2, VEGF-A, Angiopoietin-A2 as metastasis-dependent genes as well as mir-194, mir-106b, mir-126 and mir-30. It is in the MKN-45 cell line.
Methods: In this study, MKN45 cell line of gastric adenocarcinoma cells were cultured in RPMI1640 medium, then determined by MTT assay of effective LA dose and after 24 hours treatment with LA and Docetaxel changes of gene expression. The study was compared to the control group by Real Time PCR method. To evaluate the expression of the studied genes, total RNA of the treated cells was extracted at 50 mM LA and then cDNA synthesized according to the kit instructions and then analyzed by RT-PCR. In this method, U6 and beta-actin were used as housekeeping gene to measuremicroRNA and gene expression. Docetaxel was also used as a standard drug for the treatment of gastric cancer in addition to LA.
Results: Treatment of cells with LA induced changes in expression of the studied genes. Based on the results of the study, the expression of MMP-2 and Talin-2 metastasis-dependent genes was significantly increased in the treated cells compared to the untreated cells. The expression of mir-194, mir-106b, mir-126 and mir-30 was also significantly decreased in LA-treated cells. In other words, LA resulted in altered expression of genes involved in metastasis and angiogenesis