吲哚菁绿荧光成像对胰腺癌新辅助化疗疗效评估的研究

摘要

Objective To explore the imaging effectiveness of indocyanine green (ICG) in pancreatic cancer cells and curative effect evaluation of neoadjuvant chemotherapy in pancreatic cancer.Methods The basic characteristics of ICG was characterized and cell counting kit-8 (CCK-8) assay was used to observe the toxic effects of ICG and gemcitabine (GEM) in pancreatic cancer cell lines (ASPC-1 and PANC-1).After incubated with ICG for 4 hours,the ICG uptake in this two cells was observed by a high sensitivity optical imaging system (IVIS) respectively.A subcutaneous xenograft model of pancreatic cancer was established and then divided into phosphate buffer (PBS) + ICG injection group (control group) and GEM + ICG injection group (treatment group).After injection of PBS (100 mg/kg)or GEM (100 mg/kg) 24 hours,ICG (7 μg/ml) 200 μl was injected into mice by tail vein,then the fluorescence intensity in tumor tissue was observed continually by IVIS.Results ICG a1most non-toxic to cells can be excited by near-infrared light also has a good dispersion and light stability.The fluorescence intensity in both control and GEM group of ASPC-1 cells was (2.437 ± 0.039) × 107 and (0.564 ±0.327) × 107 respectively (F =13.262,P ＜ 0.05).Similarly,The fluorescence intensity in both control and GEM group of PANC-1 cells was (2.831 ± 0.029) × 107 and (0.376 ± 0.019) × 107 respectively (F =5.381,P ＜0.05).Fluorescence intensity of tumor was continuously measured by IVIS at different time points.After 4,5 and 6 hours,the tumor fluorescence intensity in GEM group was stronger than that in control group respectively (P ＜ 0.05).Conclusion ICG can be absorbed by pancreatic cancer cells and accumulate in pancreatic tumor tissues after treated with GEM.Therefore,ICG fluorescence imaging can be used to evaluate efficacy of neoadjuvant chemotherapy in Pancreatic Cancer.%目的 探索吲哚菁绿(ICG)评估胰腺癌新辅助化疗疗效的价值.方法 采用细胞计数试剂盒(CCK-8)分别观察ICG和吉西他滨(GEM)对胰腺癌细胞的毒性作用和化疗作用.ICG分别与人胰腺癌细胞株ASPC-1和PANC-1共同孵育4h后,采用高检测灵敏度的光学成像系统(IVIS)观察两种细胞摄取ICG的情况.建立胰腺癌裸鼠皮下种植瘤模型,分为磷酸盐缓冲液(PBS)+ ICG注射组和GEM+ ICG注射组,PBS(100 mg/kg)或GEM(100 mg/kg)注射24h后再次注射7μg/ml的ICG200μl,采用IVIS的BLI功能动态观察肿瘤组织内的荧光强度,并进行对比分析.结果 ICG细胞毒性小,分散性及光稳定性良好,可以由近红外光激发.IVIS检测ICG能够被胰腺癌细胞摄取并在细胞水平成像.在ASPC-1细胞中,实验组与对照组荧光强度值比较[(2.437±0.039)×107比(0.564±0.327)×107,P＜0.05)];在PANC-1细胞中,实验组和对照组荧光强度值比较[(2.831±0.029)×107比(0.376±0.019)×10 7,P＜0.05].IVIS动态测量荷瘤裸鼠肿瘤荧光强度,在第4、5、6小时,实验组肿瘤区域的荧光强度均强于对照组(P＜0.05).结论 ICG体内外均能与胰腺癌细胞结合并成像,可用于胰腺癌的分子显像和新辅助化疗疗效的评估.