Trastuzumab and folic acid functionalized gold nanoclusters as a dual-targeted radiosensitizer for megavoltage radiation therapy of human breast cancer

摘要

In the present study, the effect of functionalized gold nanoclusters (AuNCs) with trastuzumab (Herceplin (R)) and/or folic acid (FA) as a single and dual-targeted radiosensitizers for the enhancement of megavoltage radiation therapy efficacy was investigated. SK-BR3 breast cancer cells as human epidermal growth factor 2 (HER2) and folate overexpressing cell line and the murine fibroblast (L929) as a control cell line were selected. The cellular uptake was followed using inductively coupled plasma optical emission spectrometry (ICP-OES) that showed AuNCs-FA-HER uptake by SK-BR3 cells was 3 times more than the non-targeted AuNCs after 12 h incubation. MTT and clonogenic assays revealed that the viability and surviving fraction of cancer cells were significantly inhibited by treating with all AuNCs under radiation compared to treating with radiation alone. However, these effects in the dual-targeted AuNCs group (AuNCs-FA-HER) was significantly greater than non-targeted and single-targeted AuNCs groups. Also, apoptosis was evaluated using an Annexin V-FITC/propidium iodide (PI) kit in flow cytometry. All AuNCs, in combination with 4 Gy of photon beam, induced more apoptosis. By fitting the survival fraction data on the linear-quadratic model, the sensitization enhancement factor (SER) of AuNCs, AuNCs-FA, AuNCs-HER, and AuNCs-FA-HER, were obtained 1.17, 1.32, 1.48 and 1.77, respectively. SER for AuNCs-FA-HER was significantly higher than that non-targeted and single-targeted AuNCs (p-value < 0.05) that can be attributed to more internalization in the cancer cells. It was concluded that functionalized AuNCs with both folic acid and Herceptin could represent a promising strategy for increased cellular internalization that improved radiation therapy efficiency in SK-BR3 breast cancer cells.