Quantitative analysis of chemotherapeutic effects in tumors using in vivo staining and correlative histology.
Quantitative analysis of chemotherapeutic effects in tumors using in vivo staining and correlative histology.
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DOI：
10.1155/2005/791936
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发表时间：
2005
期刊：
影响因子：
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通讯作者：
Ntziachristos V
中科院分区：
文献类型：
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作者：
Choi HK;Yessayan D;Choi HJ;Schellenberger E;Bogdanov A;Josephson L;Weissleder R;Ntziachristos V
目标：使用基于膜联蛋白V-Cy 5.5探针的体内染色，在显微镜下分析肿瘤的化疗反应，并使用定量组织学分析独立评估其凋亡计数。研究方法：将对化疗敏感（CS-LLC）和耐药（CR-LLC）的刘易斯肺癌细胞植入裸鼠中并生长成肿瘤。用环磷酰胺处理小鼠并注射Cy5.5-膜联蛋白V荧光探针。使用荧光分子断层扫描进行体内成像。随后切除肿瘤并准备用于组织学。采用末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）法对组织学肿瘤切片进行细胞凋亡染色。通过TUNEL染色和相应的Cy5.5分布分析每个肿瘤的至少10个组织切片的细胞凋亡定量。结果：我们在CS-LLC肿瘤中检测到更高水平的细胞凋亡和相应更高水平的Cy5.5荧光。发现CS-LLC切片上的细胞计数率与CR-LLC的细胞计数率为0.2：1，而Cy5.5分布测量中观察到的相应面积显示CS-LLC与CR-LLC的细胞计数率为0.1.7：1。这些观察结果与CS-LLC细胞系预期的较高凋亡指数一致。结论：组织切片的定量分析显示，与CR-LLC肿瘤图像相比，CS-LLC肿瘤图像中的荧光和凋亡计数更高。这些观察结果表明，膜联蛋白V-Cy 5.5探针检测到环磷酰胺的化疗效果，并进一步证实了体内FMT测量。
Aims: To microscopically analyze the chemotherapeutic response of tumors using in vivo staining based on an annexinV-Cy5.5 probe and independently asses their apoptotic count using quantitative histological analysis. Methods: Lewis Lung Carcinomas cells, that are sensitive (CS-LLC) and resistant (CR-LLC) to chemotherapy were implanted in nude mice and grown to tumours. Mice were treated with cyclophosphamide and injected with a Cy5.5-annexinV fluorescent probe. In vivo imaging was performed using Fluorescence Molecular Tomography. Subsequently tumours were excised and prepared for histology. The histological tumour sections were stained for apoptosis using a terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay. A minimum of ten tissue sections were analyzed per tumour for apoptosis quantification by TUNEL staining and corresponding Cy5.5 distribution. Results: We detected higher levels of apoptosis and corresponding higher levels of Cy5.5 fluorescence in the CS-LLC vs. the CR-LLC tumours. The cell count rate on CS-LLC sections over CR-LLC was found to be ∼2 :1 where the corresponding area observed on Cy5.5 distribution measurements revealed a ∼1.7 :1 ratio of CS-LLC over CR-LLC. These observations are consistent with the higher apoptotic index expected from the CS-LLC cell line. Conclusions: Quantitative analysis of histological slices revealed higher fluorescence and higher apoptotic count in the CS-LLC tumour images compared to the CR-LLC tumour images. These observations demonstrate that the annexinV-Cy5.5 probe sensed the chemotherapeutic effect of cyclophospamide and further confirmed in vivo FMT measurements.