The study of TRPM5 ion channel in the line of autofacial cell in lung cancer cell lines

ABSTRACT THE STUDY OF TRPM5 ION CHANNEL IN THE LINE OF AUTOFACIAL CELL IN LUNG CANCER CELL LINES Autophagic pathways are contribute to lung cancer (LC) ethiopathogenesis. Intracellular calcium levels described as key regulator of autophagy are regulated by calcium channels. It is not known whether transient potential melastatin 5 (TRPM5) has a role in autophagic pathways. In this study, it is aimed to investigate the role of TRPM5, a calcium channel, on autophagic pathways in lung cancer A459 cell line. A459 cells were grown in RPMI-1640 medium containing 5% fetal bovine serum in a humidified incubator with 5% CO2 and 95% air at 37 °C. In A459 cell line, TRPM5 was blocked through siRNA. RNA isolation from A459 cells was conducted with Tri Reagent. mRNA expression profiles of genes related with autophagy was demonstrated by quantitative RT-PCR (qRT-PCR). Transient Receptor Potential Melastanin 5 mRNA expression level in TRPM5-siRNA group was demonstrate significiant decrease against to control group. siRNA efficiency was calculated as %83 using qRT-PCR data. In addition, qRT-PCR results demonstrated that compared with the control group there was a significant increase in autophagy related genes ATG3,ATG5, ATG10, ATG12, LC3A, LC3B, LAMP3 BECLIN1, AMBRA1, ULKE3 expression levels in TRPM5-siRNA group. Transient Receptor Potential Melastanin 5-siRNA application clearly increases the expression of genes involved in autophagic pathways. This data shows that TRPM5 suppression can activate autophagic mechanisms. In cancer it has also been determined that autophagy may show two different function in the way leading to both the survival and the death of the cell. Determining that autophagy is associated with TRPM5, by targeting TRPM5 autophagy can be inhibited or excessively increased so it may contribute to the development of cancer treatments leading to autophagic death of the cell. Key Words: Lung cancer, Authophagy, TRPM5 ion channel, Cell Culture, qRT-PCR.