evaluation of tumor regulatory genes and apoptotic pathways in the cytotoxic effect of cytochalasin h on malignant human glioma cell line (u87mg)

Objective: the aim of current study was to provide a proof-of-concept on the mechanism of plau and pcdh10 gene expressions and caspases-3, -8, and -9 activities in the apoptotic pathway after treatment of malignant human glioma cell line (u87mg) with cytochalasin h. materials and methods: in the present experimental study, we have examined cytochalasin h cytotoxic activities as a new therapeutic agent on u87mg cells in vitro for the first time. the cells were cultured and treated with 10-5-10-9 m of cytochalasin h for 24, 48 and 72 hours. the assessment of cell viability was carried out by (3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazoliumbromide (mtt) assay at 578 nm. the data are the average of three independent tests. mrna expression changes of plau and pcdh10 were then evaluated by quantitative reverse-transcriptase polymerase chain reaction (qrt-pcr). the fluorometric of caspases-3, -8, and -9 activities were carried out. the morphology changes in the u87mg cells were observed by fluorescence microscope. results: mtt assay showed that cytochalasin h (10-5 m) inhibited the u87mg cancer cells proliferation after 48 hours. analysis of qrt-pcr showed that the plau expression was significantly decreased in comparison with the control (p<0.05). the expression of pcdh10 also showed a significant increase when compared to the control (p<0.001). fluorescence microscope indicated morphological changes due to apoptosis in u87mg cancer cells, after treatment with cytochalasin h (10-5 m, 48 hours). the fluorometric evaluation of caspase-3, -8, and -9 activities showed no significant difference between the caspases and the control group. conclusion: this study shows the effect of caspase-independent pathways of the programmed cell death on the u87mg cancer cell line under cytochalasin h treatment. further studies are needed to explore the exact mechanism.