ABSTRACT
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 10-5-10-9M 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 [10-5M, 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