碳纳米管携载顺铂逆转卵巢癌耐药性的实验研究

Enhancement of Anticancer Effects of Cisplatin Carried by Single-Walled Carbon Nanotubes to Human Ovarian Carcinoma Cisplatin-Resistant Cells

目的:研究以荧光标记的大孔径碳纳米管为载体,内载顺铂,作用于顺铂耐药的卵巢癌细胞株,逆转细胞耐药性,增加顺铂对细胞的杀伤作用。方法:携带荧光标记物的碳纳米管内载顺铂后与卵巢癌耐药细胞株共培养,通过荧光显微镜检测碳纳米管载体进入细胞内的量,流式细胞仪检测载体增加顺铂对癌细胞的杀伤性能,高效液相色谱法检测载体增加细胞内顺铂含量。结果:①荧光显微镜检测结果:随着单壁碳纳米管(SWNTs)浓度(6μg/ml、12.5μg/ml、25μg/ml、50μg/ml、100μg/ml、200μg/ml)的增加,顺铂-SWNTs粒子进入卵巢癌耐药COC1/DDP细胞株的量也增加;②流式细胞仪检测:浓度高于1×10-3mg/ml时,顺铂对细胞产生了明显的凋亡效应;③高效液相色谱法检测:作用2小时后,100μg/ml和50μg/ml浓度加载体组细胞内顺铂含量[(10.1478±1.2362)×10-6ng、(9.3208±1.4302)×10-6ng]比不加载体组细胞内顺铂含量[(4.2835±0.7728)×10-6ng、(4.6852±1.0038)×10-6ng]明显增多(P<0.01)。结论:功能化的碳纳米管有望成为新的化疗药物载体,携带抗癌药物,通过增加细胞内药物浓度,逆转肿瘤耐药性,降低化疗药物的毒副反应。

Objective：To study the anticancer potential of single-walled carbon nanotubes（SWNTs） loaded with cisplatin and rhodamine in human ovarian carcinoma cisplatin-resistant cells.Methods：Cisplatin carried by SWNTs was co-cultured with human ovarian carcinoma cisplatin-resistant cells. The dosage of SWNTs vector entered into cells was observed by fluorescence microscopy The cytotoxic effect of cisplatin-loaded SWNTs on the proliferation of human ovarian carcinoma cisplatin-resistant cells was observed by flow cytometry. The determination of cisplatin in cells was observed by high performance liquid chromatography（HPLC） .Results：①The results under fluorescence microscope showed that the higher the concentration of SWNTs（6 μg/ml、12.5 μg/ml、25 μg/ml、50 μg/ml、100 μg/ml、200 μg/ml） ,the higher of cisplatin-SWNTs entering ovarian cancer Cisplatin-Resistant Cells COC1/DDP. ②Flow Cytometry showed that when the concentration was over 1×10-3 mg/ml,there was obvious apoptosis. ③ HPLC showed that there was obvious increased of cisplatin in vector plus concentration of 100 μg/ml and 50 μg/ml[（10.1478±1.2362） ×10-6 ng、（9.3208±1.4302） ×10-6 ng] than without vector group [（4.2835±0.7728） ×10-6 ng、（4.6852±1.0038） ×10-6 ng]（P0.01） 2 hours after action. Conclusions：SWNTs is to be a good vector for chemotherapy,which can carrier anticancer drugs,increase the concentration of medicine in cell,inverse tumor drug resistance,decrease the side effects of chemotherapy.