Sanad Mohammed Alonezi
PSMMC, Saudi Arabia
Title: Application of Phenotype Microarray in lung cancer after treatment with cisplatin
Biography
Biography: Sanad Mohammed Alonezi
Abstract
The phenotype microarray (PM) technique provide a high-throughput for characterization the phenotyping of cells. This technique use a redox dye, employing cell respiration (NADH production) as a universal cell based reporter. The dyes used in Biolog assays measure output of NADH production from different catabolic pathways present in the cells being tested. Figure 1. shows the layout of the carbon sources in the PM-M1 microplate. PM method applied to examine the effect of cisplatin, a chemotherapy drug, treatment on the metabolic rate in lung cancer cells produced by different substrates. The H1299 and A549 human lung cancer cell lines were tested by using standard protocols for metabolic PM mammalian cell assays. Dye reduction was calculated from A590-A750 values measured from microplate wells after 6 h from addition of MA dye. Paired t tests from univariate analysis were adjusted. In H1299 cells, there was a reduction in metabolism of dextrin, maltotriose, D-maltose, D-glucose-1-phosphate, and inosine (Figure 2) following treatment with cisplatin. In A549 cells, there was also a reduction in metabolism of substrates such as D-trehalose, D-glucose-6-phosphate and D-glucose-1-phosphate (Figure 3). The uridine, pyruvic acid and ketoglutaric acid were metabolised significantly in A549 cells after treatment with cisplatin in comparison with H1299 cells (Figure 2, 3). Adenosine substrate in the phenotype microarray plate do not appear to be useful as carbon sources in both cells. Treatment of the lung cancer cells with cisplatin produced a different effect on the H1299 cells in comparison with the A549 cells. There was more reduction in carbon metabolism in H1299 cells, but in the A549 cells the metabolism was not that strongly affected and the cells continued to produce NADH. Therefore, there may be differences in the mechanisms by which the two types of lung cancer cells respond to cisplatin treatment, which could lead to different mechanisms of cell death induced by the anticancer activity. The mechanism of necrosis and other cell death pathways may have significant difference between human lung cancer cell lines.