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Spatial and Temporal Confined Photothermolysis of Cancer Cells Mediated by Hollow Gold Nanospheres Targeted to Epidermal Growth Factor Receptors.

ABSTRACT: To date, a few studies have investigated the potential use of a short-pulsed laser in selective tumor cell destruction or its mechanism of cell killing. Computer simulation of the spatial and temporal profiles of temperature elevation after pulsed laser irradiation on an infinitesimal point source estimated that the temperature reached its highest point at ∼35 ns after a single 15 ns laser pulse. Moreover, temperature elevation was confined to a radius of sub-micrometer and returned to baseline within 100 ns. To investigate the effect of 15 ns laser pulses on A431 tumor cells, we conjugated hollow gold nanospheres (HAuNSs) to an antibody (C225) directed at the epithelial growth factor receptor. The resulting nanoparticles, C225-HAuNSs, bound to the cell membrane, internalized, and distributed throughout the cytoplasm, with some nanoparticles transported to the vicinity of the nuclear membrane. On using an optical microscope mounted to a tunable pulsed Ti:sapphire laser, rapid and extensive damage of live cancer cells was observed, whereas irradiation of A431 cells pretreated with nontargeted HAuNSs with a pulsed laser or pretreated with C225-HAuNSs with a continuous-wave laser-induced minimal cellular damage. Furthermore, after a single 15 ns laser pulse, C225-HAuNS-treated A431 cells cocultured with 3T3 fibroblasts showed signs of selective destruction. Thus, compared with a continuous-wave laser, shots of a short-pulsed laser were the most damaging to tumor cells that bound HAuNSs and generated the least heat to the surrounding environment. This mode of action by a short-pulsed laser on cancer cells (i.e., confined photothermolysis) may have potential applications in selective tumor cell destruction.

SUBMITTER: Ku G

PROVIDER: S-EPMC5981767 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Radiotherapy is preferred to chemotherapy as an adjuvant therapy for postoperative cervical cancer owing to its convenience and minimal effects on various non-targeted systems. The present study sought to investigate whether the utilization of anti-MET proto-oncogene, receptor tyrosine kinase (c-Met) antibodies conjugated to hollow gold nanospheres (anti-c-Met/HGNs) may enhance the efficiency of radiation therapy for cervical cancer. Anti-c-Met/HGNs were synthesized and confirmed to target c-Met, which was overexpressed on the cell membrane of multiple malignancies. The successful synthesis of HGNs was observed using transmission electron microscopy (TEM). Overrepresentation of c-Met in the human cervical cancer cell line CaSki was verified by immunofluorescence. The cellular uptake of HGNs was assessed using inductively coupled plasma atomic emission spectroscopy (ICP-AES). To assess the toxicity of functionalized gold nanospheres, a cell proliferation and toxicity assay was used and flow cytometry, with staining by propidium iodide (PI), was performed to study the cell cycle changes. Each experiment was conducted on three groups: Control, HGNs alone and anti-c-Met/HGNs, with each group also assessed with or without X-rays. The variation of apoptotic rate was observed by flow cytometry using a dual-staining Annexin V-fluorescein isothiocyanate/PI kit. Expression of apoptosis-associated proteins was examined by western blot analysis. TEM revealed a number of hollow spheres with cells with an average diameter of 56.25 nm and a mean wall thickness of 6.56 nm. CaSki cells were detected by inverted fluorescence microscopy via a layer of fluorescent green marker, and ICP-AES confirmed the distinct uptake of anti-c-Met/HGNs by each CaSki cell. Anti-c-Met/HGNs induced 38.7% of cells to stay in the G2/M phase, whereas the equivalent proportion in the control group was 19.8%. Compared with other groups, CaSki cells treated with anti-c-Met/HGNs and 5 Gy X-ray radiation exhibited a higher apoptosis rate (16.92%) and a higher early apoptotic rate (12.30%) compared with cells under other conditions (control+0 Gy: 3.16 and 1.69%; HGN+0 Gy: 3.98 and 1.94%; anti-c-Met/HGN+0 Gy: 3,47 and 1.85%; control+5 Gy: 5.35 and 3.66%; HGN+5 Gy: 7.91 and 4.06%). The anti-c-Met/HGN X-ray-treated group also evidently overexpressed caspase-3 and BCL2 associated X, apoptosis regulator. Anti-c-Met/HGN may, therefore, aid the sensitivity of radiation therapy in cervical cancer.

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Spatial and Temporal Confined Photothermolysis of Cancer Cells Mediated by Hollow Gold Nanospheres Targeted to Epidermal Growth Factor Receptors.

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