摘要
目的:探究光敏剂DCFP-Cl介导的光动力治疗(photodynamic therapy,PDT)对胆管癌细胞系的生物安全性和有效性。方法:人肝胆管癌细胞(RBE)与DCFP-Cl共培养,观察DCFP-Cl在细胞内的荧光成像潜力,引入Mito-Tracker Green观察DCFP-Cl的细胞器靶向性;使用Calcein AM、DCFH-DA试剂盒检测PDT后细胞活性及细胞内单态氧产生情况;使用CCK-8法定量评估DCFP-Cl对细胞的暗毒性、光动力效率;通过FACS法初步探索DCFP-Cl介导光动力的作用机制;构建荷瘤裸鼠模型,进行动态活体荧光成像,观察DCFP-Cl体内荧光成像能力,通过分组治疗,观察DCFP-Cl对活体肿瘤的抑制作用及生物安全性。结果:在RBE细胞内可见DCFP-Cl的红色荧光与Mito-Tracker Green的绿色荧光重叠,皮尔森相关系数为0.78±0.07,呈强相关;接受PDT组,细胞活力检测实验中未见明显绿色荧光,活性氧实验中显示出明显的绿色荧光;在光照下,随着DCFP-Cl浓度梯度增加RBE细胞存活率逐渐降低,IC50为7.12μmol/L;通过FACS分析,DCFP-Cl介导的PDT诱导细胞死亡的方式主要为细胞凋亡,总体凋亡率随辐照时间延长而增加(t=5.726,P<0.05);荷瘤裸鼠通过尾静脉注射DCFP-Cl,2 h后DCFP-Cl开始在肿瘤区域积聚,以8 h聚集效果最佳;分组治疗中,PDT组肿瘤的大小明显小于其他三组(F=26.82,P<0.001),其HE染色中肿瘤细胞膜不完整,TUNEL染色中可见明显的细胞凋亡,且该组裸鼠主要脏器均未受到损害。结论:DCFP-Cl介导的PDT在胆管癌的体内、外实验中均展现了良好的生物低暗毒性、稳定的荧光成像能力和优异的抗肿瘤活性。
Objective:To evaluate the biosafety and efficacy of DCFP-Cl-mediated photodynamic therapy(PDT)in cholangiocarcinoma cell lines.Methods:Co-culturing human liver cholangiocarcinoma cells(RBE)with DCFP-Cl,we assessed the potential of DCFP-Cl for intracellular fluorescence imaging.We introduced Mito-Tracker Green to investigate the organelle specificity of DCFP-Cl.Cell viability and the generation of intracellular singlet oxygen post-photodynamic therapy were determined using Calcein AM and DCFH-DA assays.The CCK-8 assay quantitatively evaluated the cytotoxicity and photodynamic efficiency of DCFP-Cl.Furthermore,the FACS method provided preliminary insights into the mechanistic aspects of DCFP-Cl-mediated photodynamic therapy.For in vivo analysis,we established a nude mouse model bearing tumors for dynamic fluorescence imaging to monitor the fluorescence,which strongly overlapped with the green fluorescence of Mito-Tracker Green(Pearson correlation coefficient:0.78±0.07).Post-photodynamic treatment,significant green fluorescence indicating reactive oxygen species was observed,contrasting with the absence of such fluorescence in cell viability assays.As the DCFP-Cl concentration increased,RBE cell survival progressively decreased,with an IC50 value of 7.12μmol/L.FACS analysis indicated that DCFP-Cl-mediated PDT predominantly triggered apoptosis,with increasing apoptotic rates correlating with extended irradiation times(t=5.726,P<0.05).In vivo studies involved administering DCFP-Cl to tumor-bearing nude mice via tail vein injection,leading to optimal tumor accumulation at 8 hours post-injection.Compared to control groups,the photodynamic therapy group exhibited significantly reduced tumor sizes(F=26.82,P<0.001),with histological analysis revealing disrupted tumor cell membranes and marked apoptosis.Notably,no significant organ damage was observed in these mice.Conclusion:DCFP-Cl-mediated photodynamic therapy has demonstrated promising biosafety,stable fluorescence imaging,and potent anti-tumor efficacy in both in vit
作者
吴琳
舒禹铭
郭航成
田野
宋锋玲
郑文恒
WU Lin;SHU Yuming;GUO Hangcheng;TIAN Ye;SONG Fengling;ZHENG Wenheng(Department of Interventional Therapy,Cancer Hospital of Dalian University of Technology(Liaoning Cancer Hospital&Institute),Liaoning Shenyang 110042,China;China Medical University,Liaoning Shenyang 110122,China;Colorectal Surgery,Cancer Hospital of Dalian University of Technology(Liaoning Cancer Hospital&Institute),Liaoning Shenyang 110042,China;Institute of Frontier and Interdisciplinary Science,Shandong University,Shandong Qingdao 266237,China)
出处
《现代肿瘤医学》
CAS
2024年第11期1955-1963,共9页
Journal of Modern Oncology
基金
中央高校基本科研业务费资助项目(编号:LD202016)。
关键词
胆管癌
荧光成像
光动力治疗
光敏剂
线粒体
cholangiocarcinoma
fluorescence imaging
photodynamic therapy
photosensitizer
mitochondria
