Abstract 121P
Background
Glioblastoma is a highly aggressive brain tumor with limited treatment options and high recurrence rates due to the challenges in targeting infiltrative cells. Literarture suggests that calcite, a calcium carbonate crystal, exhibits anticancer properties, including the inhibition of lung cancer cell propagation, likely through alkalization effects. This study examines the potential of calcite as a cytotoxic agent against glioblastoma, using the A172 human glioblastoma cell line.
Methods
The dose-response of calcite on A172 cells was assessed through MTT Assay and Hoechst-Propidium Iodide (PI) staining. Morphological changes were examined using Scanning Electron Microscopy and fluorescence microscopy while proliferation was evaluated using Ki-67 immunofluorescence. Clonogenic and scratch wound assays measured colony formation and cell migration. Apoptosis was determined by Acridine Orange-Ethidium Bromide staining and primary glial cell viability was determined by Hoechst- Fluorescein diacetate (FDA)-PI staining.
Results
Calcite particles inhibited cell growth in a dose- and time-dependent manner. One day post-treatement, growth inhibition ranged from 2.37% to 32% (0.01–5 mg/mL), increasing to 39% (1 mg/mL) and 59% (5 mg/mL) at day 5. Calcite teatment induced changes in cell circularity and solidity. After 24hrs of treatment, Ki-67 expression was reduced to 54% and 47% at 1mg/mL and 5mg/mL, respectively, compared to untreated control (72%). Calcite (1 and 5 mg/mL) suppressed the colony formation of A172 cells by 64% and 77 % and inhibited cell migration by 47% and 64%, respectively. Apoptosis analysis showed an 18-fold increase in apoptotic cells at 5mg/mL and a 7-fold increase at 1mg/mL compared to untreated control. By contrast, calcite had minimal impact on postnatal rat hippocampal glial cells, resulting in 9.41% and 23.43% total cell reduction at 1 mg/mL and 5 mg/mL, respectively.
Conclusions
Calcite inhibits the growth and proliferation of A172 cells by reducing viability, migration and colony formation while increasing apoptosis. Importantly, it exhibits selective toxicity towards glioblastoma cells over normal glial cells. These results suggest its potential as a targeted therapeutic agent for anti-glioblastoma treatments.
Clinical trial identification
Editorial acknowledgement
Legal entity responsible for the study
Danny Baranes.
Funding
LifeForce Labs, Israel.
Disclosure
All authors have declared no conflicts of interest.