叶酸修饰的克班宁纳米粒联合超声辐照对M109细胞的体内外靶向作用及抗肿瘤机制-中国药房

叶酸修饰的克班宁纳米粒联合超声辐照对M109细胞的体内外靶向作用及抗肿瘤机制

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篇名：	叶酸修饰的克班宁纳米粒联合超声辐照对M109细胞的体内外靶向作用及抗肿瘤机制
TITLE：	Targeting effect and anti-tumor mechanism of folic acid-modified crebanine nanoparticles combined with ultra-sound irradiation on M109 cells in vitro and in vivo
摘要：	目的考察叶酸修饰的克班宁纳米粒（FA-Cre@PEG-PLGANPs，以下简称“NPs”）联合超声辐照对M109细胞的体内外靶向作用，初步探讨其抗肿瘤机制。方法采用CCK-8法检测NPs联合超声辐照对M109细胞的增殖抑制作用，筛选最佳超声时间。以肺癌A549细胞为对照，通过游离叶酸阻断实验和细胞摄取实验，评估NPs联合超声辐照对M109细胞的叶酸靶向性。利用细胞划痕实验、Transwell小室实验和流式细胞术，检测给药后1h联合超声辐照下，NPs对M109细胞迁移、侵袭、凋亡、周期及细胞内活性氧（ROS）水平的影响；使用荧光倒置显微镜观察线粒体膜电位（MMP）变化。构建M109细胞的小鼠皮下瘤模型，通过小动物活体成像技术考察NPs联合超声辐照的体内肿瘤靶向性。结果NPs联合超声辐照能明显抑制M109细胞的增殖，最佳超声时间点为给药后1h。游离叶酸可以拮抗NPs对M109细胞的增殖抑制作用，联合超声辐照能部分逆转这一拮抗作用。与A549细胞比较，M109细胞对NPs的摄取能力更强（P＜0.01），超声辐照可以促进细胞摄取。NPs联合超声辐照可以抑制M109细胞的迁移与侵袭，并将细胞阻滞于G0/G1和G2/M期。与对照组比较，NPs不同质量浓度（100、200、300μg/mL）组M109细胞的凋亡率、ROS水平均显著升高（P＜0.01），MMP均显著降低（P＜0.01）。与不超声组比较，给药后0h超声组小鼠肿瘤部位的荧光强度及肿瘤靶向指数均显著增加（P＜0.05或P＜0.01）。结论NPs联合超声辐照对M109细胞具有较强的体内外靶向性，其抗肿瘤机制包括抑制细胞迁移与侵袭、阻滞细胞周期、诱导细胞凋亡等。
ABSTRACT：	OBJECTIVE To investigate the targeting effect of folic acid-modified crebanine nanoparticles （FA-Cre@PEG- PLGA NPs， hereinafter referred to as “NPs”） combined with ultrasound irradiation on M109 cells in vitro and in vivo after administration， and explore the anti-tumor mechanism. METHODS CCK-8 assay was used to detect the inhibitory effect of NPs combined with ultrasound irradiation on the proliferation of M109 cells， and the best ultrasound time was selected. Using human lung cancer A549 cells as a control， the targeting of NPs combined with ultrasound irradiation to M109 cells was evaluated by free folic acid blocking assay and cell uptake assay. The effects of NPs combined with ultrasound irradiation on the migration， invasion， apoptosis， cell cycle and reactive oxygen species （ROS） levels of M109 cells were detected by cell scratch test， Transwell chamber test and flow cytometry at 1 h after 958401536@qq.com administration； the changes of mitochondrial membrane potential （MMP） were observed by fluorescence inverted microscope. A mouse subcutaneous tumor model of M109 cells was constructed， and the in vivo tumor targeting of NPs combined with ultrasound irradiation was investigated by small animal in vivo imaging technology. RESULTS NPs combined with ultrasound irradiation could significantly inhibit the proliferation of M109 cells， and the optimal ultrasound time was 1 h after administration. The free folic acid could antagonize the inhibitory effect of NPs on the proliferation of M109 cells， and combined with ultrasound irradiation could partially reverse this antagonism. Compared with A549 cells， the uptake rate of NPs in M109 cells was significantly higher （P＜0.01）， and ultrasound irradiation could promote cellular uptake. NPs combined with ultrasound irradiation could inhibit the migration and invasion of M109 cells and block the cell cycle in the G0/G1 and G2/M phases. Compared with control group， the apoptosis rate of M109 cells and ROS level were increased significantly （P＜0.01）， while the MMP decreased significantly （P＜0.01） in the different concentration （100， 200， 300 μg/mL） groups of M109 cells. Compared with the mice in non-ultrasound group， the fluorescence intensity and tumor-targeting index of the tumor site in the 0 h ultrasound group were significantly enhanced （P＜0.05 or P＜0.01）. CONCLUSIONS NPs combined with ultrasound irradiation have a strong targeting effect on M109 cells in vitro and in vivo， the anti-tumor mechanism includes inhibiting cell migration and invasion， blocking cell cycle， and inducing apoptosis.
期刊：	2025年第36卷第14期
作者：	张海亮；赵笑雨；梅佳华；潘蕊；唐钧泽；喻锟；薛蕊；李晓飞；程欣
AUTHORS：	ZHANG Hailiang， ZHAO Xiaoyu，MEI Jiahua，PAN Rui，TANG Junze，YU Kun，XUE Rui，LI Xiaofei，CHENG Xin
关键字：	叶酸修饰纳米粒；超声辐照；M109细胞；肿瘤靶向性；抗肿瘤机制
KEYWORDS：	folic acid-modified nanoparticles； ultrasonic irradiation； M109 cells； tumor targeting； anti-tumor mechanism
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