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Womens Health

Women's Health - Poster Session D

Poster L20 - Novel Method of Delivering Low Dose Ionizing Radiation to Breast Epithelial Cells

Friday, October 25, 2024
3:30 PM – 4:30 PM EST
Location: Exhibit Hall E, F & G

Introduction:  Ionizing radiation is a common treatment method for various types of cancer. This method typically consists of high doses of radiation in order to treat the cancer. While the effects of high doses of ionizing radiation are well understood, the effects on cells of low energy ionizing radiation (LER) is less understood. While these absorbed doses are much larger in the tumor, we are regularly experiencing LER through man-made and natural sources. Examples of naturally occurring sources of LER include: background radiation from space as well as naturally occurring materials such as radon gas that can be found in basements. As well as, man made sources include diagnostic x-rays and power plants. Our project focuses on using our optimized x-ray setup to irradiate breast epithelial cells, MCF-12s to determine the effects on the proliferation.

Materials and

Methods:  For these experiments, breast epithelial cells, MCF-12s, were used. Cells were cultured in DMEM-F12 with 10% Fetal Bovine Serum (FBS), 0.1% Antibiotic-Antimycotic (AA), and 10 μg/ml of Bovine insulin. The cells were cultured under standard culture conditions (5% CO2, 37℃) until ready for experimentation. In order to create varying doses, we used two different Al filter thicknesses allowing us to have our control group, low dose group, and high dose group. The cells were irradiated in our modified irradiator setup at 10kV and 4μA. Immediately after irradiation, we performed a CellTiter 96® AQueous Cell Proliferation Assay on one of the samples from each group and imaged using Cytation 5 plate reader. The other samples were incubated overnight. This assay was run 24hrs and 48hrs after irradiation as well.

Results, Conclusions, and Discussions: We successfully cultured the MCF-12s as described and ran the proliferation assay at the three different time points. Based on the results seen in the figure, we can conclude that immediately after irradiation, the proliferation was higher in the thin filter group. In the thin filter group, there was a clear decrease in the proliferation of the cells overtime. In the control and thick filter group, we saw an increase in the proliferation at the 24hr time point, but a decrease at the 48hr time point. We found that in all three groups the proliferation levels were about the same at the 48hr time point. There was a significant difference in the proliferation between the control and thin filter group at the 0hr time point (p=0.0094). There was no significant difference between the control and thin filter group at the 24hr time point (p=0.68).
Based on the results of the proliferation assay, we are able to conclude that MCF-12s do not seem to be a radiosensitive cell type compared to other cell lines such as 3T3s. This work expands on previous work that demonstrated that radiation has a significant effect on the proliferation of some cell types. Our work contributes to the understanding of radioprotection and cancer treatment methods. Our modified irradiator setup will allow us to use more diverse doses and energies that will contribute to the understanding of radiation at the cellular level.
Future work should include cancer cell lines, such as MCF-7s in order to determine the effects of the low energy radiation on cancer cells. We would also like to run morphology or immunohistology staining before and after irradiation to determine if the radiation had an effect on the morphology. An ROS assay should be used in future experimentation to determine if the radiation is causing an increase in free radicals as this could lead to varying levels of proliferation. Additional time points for the proliferation assay should be taken such as 12hrs and 36hrs as there was a clear change between the 0hr and 24hr time points.

Acknowledgements (Optional):  We would like to thank the members of the multiscale bioelectromechanics lab at Clemson University. This material is based upon work supported by the National Science Foundation under Grant No. 2349368.