CAJ | 학술논문

目的：构建新型曲妥珠单抗－纳米金生物探针（herceptin－GNPs），探寻纳米靶向治疗乳腺癌新方法。方法：采用柠檬酸三钠还原法制备金纳米粒子（GNPs），通过静电吸附作用偶联曲妥珠单抗（herceptin）合成herceptin－GNPs；MTT法检测GNPs、herceptin、herceptin－GNPs对HER2阳性乳腺癌BT474细胞体外增殖抑制作用；流式细胞术检测BT474细胞凋亡率和周期变化；蛋白质印迹技术检测p－AKT和Bcl－2蛋白表达。结果：GNPs水溶液澄清透亮，静电吸附成功制备herceptin－GNPs探针。 GNPs浓度在100μg? ml－1及以下未显示明显抑制BT474细胞增殖作用。不同浓度herceptin和herceptin－GNPs对BT474细胞生长具有剂量依赖的抑制作用；与herceptin 相比， herceptin－GNPs抑制作用更强。流式细胞术检测显示herceptin、herceptin－GNPs均诱导 BT474细胞凋亡，阻滞细胞于 G0／G1期，且 herceptin－GNPs 作用强于 herceptin。 herceptin、herceptin－GNPs均可下调抑凋亡蛋白 Bcl－2表达，降低AKT的磷酸化水平，且 herceptin－GNPs 作用强于herceptin。结论：本研究合成的herceptin－GNPs具有良好的生物相容性，较herceptin抑制乳腺癌BT474细胞生长作用更强，可减少herceptin给药剂量。 herceptin－GNPs可能为HER2过表达乳腺癌的靶向治疗提供新方法。
목적：구건신형곡타주단항－납미금생물탐침（herceptin－GNPs），탐심납미파향치료유선암신방법。방법：채용저몽산삼납환원법제비금납미입자（GNPs），통과정전흡부작용우련곡타주단항（herceptin）합성herceptin－GNPs；MTT법검측GNPs、herceptin、herceptin－GNPs대HER2양성유선암BT474세포체외증식억제작용；류식세포술검측BT474세포조망솔화주기변화；단백질인적기술검측p－AKT화Bcl－2단백표체。결과：GNPs수용액징청투량，정전흡부성공제비herceptin－GNPs탐침。 GNPs농도재100μg? ml－1급이하미현시명현억제BT474세포증식작용。불동농도herceptin화herceptin－GNPs대BT474세포생장구유제량의뢰적억제작용；여herceptin 상비， herceptin－GNPs억제작용경강。류식세포술검측현시herceptin、herceptin－GNPs균유도 BT474세포조망，조체세포우 G0／G1기，차 herceptin－GNPs 작용강우 herceptin。 herceptin、herceptin－GNPs균가하조억조망단백 Bcl－2표체，강저AKT적린산화수평，차 herceptin－GNPs 작용강우herceptin。결론：본연구합성적herceptin－GNPs구유량호적생물상용성，교herceptin억제유선암BT474세포생장작용경강，가감소herceptin급약제량。 herceptin－GNPs가능위HER2과표체유선암적파향치료제공신방법。
Objective:To construct an innovative biocompatible nanoprobe herceptin-gold nanoparticles( GNPs) , and to explore new targeted therapies in breast cancer.Methods: GNPs were synthesized by the classic citrate reduction method and the herceptin conjugated to GNPs by electrostatic adsorption. Setting 5 concentration gradients of GNPs from 6.25μg? ml -1 to 100μg? ml-1 , herceptin and herceptin-GNPs from 0.625μg? ml -1 to 10 μg? ml -1 along with their control group to detect the cytotoxicity by MTT assay.The apoptosis, cell cycle changes were detected by flow cytometry and the expressions of p-AKT and Bcl-2 protein were detected by Western Blot, respectively.Results: The GNPs aqueous solution obtained were fuchsia and clear.The GNPs showed no cytotoxicity under the designed concentration gradients, which displayed good biocompatibility.Herceptin and herceptin-GNPs both could suppress cell growth, induce apoptosis and block cell proliferation in G0/G1 phase;the latter had better effect.Moreover, both herceptin and herceptin-GNPs could inhibit the expressions of p-AKT and Bcl-2 protein, and the latter had better inhibitory effect.Conclusion: The herceptin-GNPs are biocompatible. Compared with herceptin, the herceptin-GNPs have better inhibitory effect on HER2 overexpressing breast cancer cell proliferation, which can reduce the dosage of herceptin.It would provide a nanoparticle-based targeted delivery system for the treatment of HER2 overexpressing breast cancer.