Determination of gremlin-1 expression levels in glioblastoma cell processed with lipopolysaccharide and temozolomide

Glioblastoma (GB) is the most known maling brain tumor. Recent studies have shown gremlin 1 as a potential therapeutic target for certain types of cancer. Gremlin 1 is thought to be expressed in tumor tissues by cancer-related stromal cells, thereby contributing to tumor growth and invasion regulated by microenvironment, and gremlin1 expression has been associated with tumor development. Temozolomide (TMZ) is often used in chemotherapy treatment of Glioblastoma Multiforme and is the only alkylating agent with an anti-tumor effect. Lipopolysaccharide is a bacterial cell wall element that stimulates tumor cells and has an anti-tumor effect on gliomas. In this experimental study; U-118 human glioblastoma multiforme cell line was replicated in cell culture medium, and cells were removed from the flask with trypsin-EDTA on the day when approximately 95% density was observed. The cell suspension was prepared and the cells were added to 60 mm2 petri dishes and they were allowed to multiply. The cells reaching the desired density were applied at 200 µM, 100 µM, 50 µM, 25 µM concentration ranges, TMZ and LPS at 10 µg / ml, 1 µg / ml, 0.1 µg / ml, 0.01 µg / ml concentration ranges. Protein isolation was performed immediately after the incubation times of 24 hours and 48 hours. Gremlin 1 protein levels were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. In our study, we showed an increase in gremlin-1 expressions in 24-hour 25 and 50 µM concentrations of TMZ-treated glioblastoma cell lines, and a decrease in Gremlin-1 expression in glioblastoma cells that received LPS at a concentration of 0.1 µg / ml for 24 hours. These findings indicate that gremlin-1 levels play an important role in glioblastoma. Considering the frequency of resistance development and shortness of survival in glioblastoma tumors, we believe that investigating the effectiveness of gremlin-1 can be quite significant.