Abstract
Background: This study investigated the effectiveness and underpinning mechanisms of radiosensitization using octaarginine (R8)-modi ed gold nanoparticle–poly(ethylene glycol) (GNP-PEG-R8) in colorectal cancer cell line LS180 to megavoltage radiotherapy in vitro.
Method: In-house synthesized GNP-PEG was characterized by transmission electron micro- scopy, dynamic light scattering, ultraviolet–visible spectrophotometry, and X-ray photoelectron spectroscopy. Inductively coupled plasma mass spectroscopy was used to quantify internaliza- tion. Direct cytotoxicity was established using the Cell Counting Kit-8, while radiosensitivity was determined using the gold standard in vitro clonogenic assay. Cell-cycle distribution, apoptosis, reactive oxygen species (ROS), and mitochondrial membrane potential (MMP) were analyzed by ow cytometry, further exploring the key mechanisms driving GNP-PEG-R8 radiosensitization.
Results: The core GNP diameter was 6.3±1.1 nm (mean±SD). Following functionalization, the hydrodynamic diameter increased to 19.7±2.8 nm and 27.8±1.8 nm for GNP-PEG and GNP- PEG-R8, with respective surface plasmon resonance peaks of 515 nm and 525 nm. Furthermore, incorporation of the R8 signi cantly increased nanoparticle internalization compared to GNP- PEG (p,0.001) over a 1 h treatment period. Functionalized GNPs confer little cytotoxicity below 400 nM. In clonogenic assays, radiation combined with GNP-PEG-R8 induced a sig- ni cant reduction in colony formation compared with radiation alone, generating a sensitizer enhancement ratio of 1.59. Furthermore, GNP-PEG-R8 plus radiation predominantly induced cell-cycle arrest in the G2/M phase, increasing G2/M stalling by an additional 10% over GNP- PEG, markedly promoting apoptosis ( p,0.001). Finally, ROS levels and alterations in MMP were investigated, indicating a highly signi cant ( p,0.001) change in both parameters following the combined treatment of GNP-PEG-R8 and radiation over radiation alone.
Conclusion: R8-modi ed GNPs were ef ciently internalized by LS180 cells, exhibiting minimal cytotoxicity. This yielded signi cant radiosensitization in response to megavoltage radiation. GNP-PEG-R8 may enhance radiosensitivity by arresting cell cycle and inducing apoptosis, with elevated ROS identi ed as the likely initiator.
Method: In-house synthesized GNP-PEG was characterized by transmission electron micro- scopy, dynamic light scattering, ultraviolet–visible spectrophotometry, and X-ray photoelectron spectroscopy. Inductively coupled plasma mass spectroscopy was used to quantify internaliza- tion. Direct cytotoxicity was established using the Cell Counting Kit-8, while radiosensitivity was determined using the gold standard in vitro clonogenic assay. Cell-cycle distribution, apoptosis, reactive oxygen species (ROS), and mitochondrial membrane potential (MMP) were analyzed by ow cytometry, further exploring the key mechanisms driving GNP-PEG-R8 radiosensitization.
Results: The core GNP diameter was 6.3±1.1 nm (mean±SD). Following functionalization, the hydrodynamic diameter increased to 19.7±2.8 nm and 27.8±1.8 nm for GNP-PEG and GNP- PEG-R8, with respective surface plasmon resonance peaks of 515 nm and 525 nm. Furthermore, incorporation of the R8 signi cantly increased nanoparticle internalization compared to GNP- PEG (p,0.001) over a 1 h treatment period. Functionalized GNPs confer little cytotoxicity below 400 nM. In clonogenic assays, radiation combined with GNP-PEG-R8 induced a sig- ni cant reduction in colony formation compared with radiation alone, generating a sensitizer enhancement ratio of 1.59. Furthermore, GNP-PEG-R8 plus radiation predominantly induced cell-cycle arrest in the G2/M phase, increasing G2/M stalling by an additional 10% over GNP- PEG, markedly promoting apoptosis ( p,0.001). Finally, ROS levels and alterations in MMP were investigated, indicating a highly signi cant ( p,0.001) change in both parameters following the combined treatment of GNP-PEG-R8 and radiation over radiation alone.
Conclusion: R8-modi ed GNPs were ef ciently internalized by LS180 cells, exhibiting minimal cytotoxicity. This yielded signi cant radiosensitization in response to megavoltage radiation. GNP-PEG-R8 may enhance radiosensitivity by arresting cell cycle and inducing apoptosis, with elevated ROS identi ed as the likely initiator.
| Original language | English |
|---|---|
| Pages (from-to) | 3541-3552 |
| Journal | International Journal of Nanomedicine |
| Volume | 13 |
| DOIs | |
| Publication status | Published - 19 Jun 2018 |