In this study, zeolite X synthesised from coal fly ash (CFA) was examined for its anti-proliferative effects on human breast cancer MCF-7 cells by in vitro studies and allied probable molecular mechanism were also assessed. MTT assay exposed that zeolite X showed a concentration-dependent inhibitory result on the proliferation of MCF-7 cells and caused early apoptotic death and does not induce cytotoxicity in non-cancerous cells (MCF-12A). Flow cytometric study specified the accrual of cells at S phase suggest induction of apoptosis which was avowed through fluorescence and confocal microscopy following annexin V-FITC/propidium iodide (PI). MCF-7 cells treated by 10, 15 and 20 µg/ml concentrations of zeolite X showed internucleosomal DNA fragments in ladder form, thereby indicates cell death is associated with apoptosis. Mechanistically, our data support the induction of apoptosis through the activation of mitochondrial dependent pathway as indicated by an up-regulation of Bax and downregulation of Bcl-2 ratio, the expulsion of cytochrome c from the mitochondria to the cytosol and cleavage of pro-caspase-3 into activation of caspase-3 in MCF-7 cells. Based on these outcomes zeolite X induced early apoptosis and strongly provided experimental evidence for the usage of zeolite X as an essential therapeutic agent in the prevention and therapy of human breast cancer.
The synthesised zeolite X from coal fly ash showed significant cytotoxic activity in contradiction of HeLa cells (cervical cancer) in a concentration-dependent way at concentrations ranges from 200 µg to 0.781 µg/ml as shown by MTT assay and failed to cause cytotoxic effect in normal cells (Gh239). Cell cycle analysis exposed that zeolite X (10 and 15 µg/ml) endorses cell growth inhibition by inducing G2/M phase arrest in HeLa cells as observed using flow cytometry. The confocal microscopic results depicted increased early apoptotic related changes in HeLa cell lines induced by zeolite X at a dosage of 10, 15 and 20 µg/ml. Zeolite X at a dosage of 10, 15 and 20 µg/ml in HeLa cells showed fragmentation of DNA by ladder pattern thereby indicates that cell death is related with apoptosis. By the increase of Bax/Bcl-2 ratio, zeolite X leads to the caspase-3 and caspase-9 activation and allow the cells to enter apoptosis. These collective results evidently showed that the influence of mitochondria-mediated signalling pathway in zeolite X induced apoptosis and intensely delivered investigational suggestion for the use of zeolite X as a significant curative agent in the preclusion and therapy of human cervical carcinoma.
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