To conduct an in vivo optical imaging analysis of the biodistribution of antibody Rituximab in lymphoma tumor-bearing nude mice.
Laser scanning confocal microscope and flow cytometry were employed to determine the affinity of FITC-labeled Rituximab (FITC-Rituximab) with human lymphoma Raji cells. And the in vivo optical imaging system was used to analyze the biodistribution of FITC-Rituximab in lymphoma-transplanted xenograft nude mice.
The results of flow cytometry and laser scanning confocal microscope demonstrated that FITC-Rituximab had remarkable affinity with lymphoma Raji cells and was mainly bound at cell membrane. The results of in vivo imaging analysis suggested that FITC-Rituximab could specifically accumulated at peritumor tissue less than 1 h, then penetrated into the interior of tumor and concentrated in 3-4 h. And the specific concentration of FITC-Rituximab could still been observed more than 8-10 h whereas there was no apparent fluorescence at other tissues. Furthermore, the results observed from a two-flank tumor xenograft model showed that FITC-Rituximab possessed specific binding affinity for CD20-overexpressed lymphoma.
The in vivo optical imaging system can accurately monitor the distribution of FITC-Rituximab in tumor-bearing nude mice. And this technique has a reference value and significance for a real-time analysis of tumor-targeting capability of antibody drugs.
[Show abstract][Hide abstract] ABSTRACT: Human xenograft mouse models, which have been used in cancer research for over a century, provided significant advances for our understanding of this multifaceted family of diseases. Orthotopic transplantation tumor models are emerging as the preference for cancer research due to the increasing clinical relevance over subcutaneous mouse models. In this study, a stable luciferase-expressed Capan-2 cell line was constructed and the expression of luciferase was tested. The results showed that the luminorescence intensity of Capan-2Luc cells was associated with the number of cells and the minimal detectable cell population was 600 cells/well. We established an orthotopic transplantation model of pancreatic cancer using Capan-2Luc cell line in athymic mice and investigated the inhibitory effects of gemcitabine (Gem) in vitro and in vivo. Optical imaging system was applied to evaluate the tumor growth of orthotopic transplantation model in vivo. The results suggested that the orthotopic transplantation model of pancreatic cancer was well established and the luminorescence intensity of Gem-treated group was markedly lower than that of control group with an inhibitory rate of 56.8% (P<0.001). Our orthotopic transplantation model of pancreatic cancer and real-time imaging observation method established in this study could be an ideal model and a useful tool for therapeutic approaches for pancreatic cancers.
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