RESUMO
Background and purpose: Renal cell carcinoma is the most common form of kidney cancer, characterized by lack of early symptoms and high malignancy. This study aimed to establish orthotopic nude mice models of human renal cell carcinoma with high success rate and good repeatability. Methods: The four types of methods which were adopted to establish the orthotopic models of renal cell carcinoma were orthotopic injection of 786-0 and ACHN cell suspensions, orthotopic injection of primary cell suspensions obtained from the subcutaneous tumor tissues, renal subcutis orthotopic implantation into renal capsule and surgical subcutis orthotopic implantation into renal fascia. To gain insights into the tumorigenicity and the growth of transplantation tumors, the imageological examination (PET/CT), histological examination (H-E staining, immunohistochemistry staining) and biochemical analysis of blood were carried out. Results: In terms of the subcutaneous transplantation of human renal cell carcinoma models in nude mice, tumorigenic rate of ACHN cells (90%) was higher than that of 786-0 cells (30%). The tumorigenic incidences of 786-0 cell suspensions orthotopic injection, ACHN cell suspensions orthotopic injection, ACHN subcutis cellular suspensions orthotopic injection, ACHN subcutis orthotopic implantation into renal capsule and renal fascia were 33%, 80%, 90%, 100% and 20%, respectively. ACHN subcutis orthotopic implantation into renal capsule was the most effective approach. Imageological and histological results accorded with poorly differentiated renal cell carcinoma. Conclusion: Four orthotopic nude mice models of human renal cell carcinoma were successfully established. Among these methods, ACHN subcutis orthotopic implantation into renal capsule is the most effective approach, which provides an ideal model for the research on biological behavior of human renal cell carcinoma and its treatment.
RESUMO
BACKGROUND:Studies have testified that nano-ultrasound contrast agents have a strong permeability, making it possible to image the targeted tissues outside blood vessels and overcome the limitation that micron contrast agents are only available for the blood pool imaging. OBJECTIVE:To construct the folate-modified nanoparticles targeting breast cancer as ultrasound contrast agents, as wel as to observe their ability to specifical y bind to cel s and imaging effect in vitro. METHODS:Both contrast agents, pegylated lactic acid-glycolic acid copolymer wrapping liquid fluorocarbon formed nanoparticles (mPP/PFOB) and folate modified pegylated lactic acid-glycolic acid wrapping liquid fluorocarbon formed nanoparticles (mPPF/PFOB), were constructed by phacoemulsification-evaporation method. (1)Biocompatibility detection:HFF-1 and MCF-7 cel s in the logarithmic phase were cultivated with various concentrations (0, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2 and 1 g/L) of mPP/PFOB or mPPF/PFOB for 24 hours respectively, and then the cel viability was measured. (2)Targeting ability detection in vitro:HFF-1 and MCF-7 cel s in the logarithmic phase were divided into three groups. Cy5-labled mPP/PFOB and mPPF/PFOB were added into groups A and B, respectively;the cel s in group C were pretreated with folate for 2 hours, and sequential y Cy5-labled mPPF/PFOB was added into group C. Fluorescence intensity was detected by flow cytometry after 0.5 hours of culture. The distribution of contrast agents in cel s was observed using confocal microscopy after 20 minutes of culture. (3)Ultrasound imaging in vitro:there were three groups:saline was as group A;the suspension of saline and mPPF/PFOB nanoparticles was prepared as group B;MCF-7 cel s were resuspended with the mixture of saline and mPPF/PFOB nanoparticles to prepare the suspension of nanoparticles and cel s as group C. In each group, the suspension was added into latex gloves, that were then tightened and immersed in water. Final y, the ultrasound was use to detect the ultrasound imaging effect in vitro. RESULTS AND CONCLUSION:Neither nanoparticles were with significant cytotoxicity. The flow cytometry showed that the mean fluorescence intensity in MCF-7 cel s of group B was significantly higher than that of groups A and C. But there were no significant differences in the mean fluorescence intensity in HFF-1 cel s among the three groups. It was observed that mPPF/PFOB mainly gathered around the MCF-7 cel membrane, while mPP/PFOB randomly distributed in the cytoplasm. After mPPF/PFOB binding to MCF-7 cel s, they could enhance ultrasound echo in vitro. These findings indicate that the targeted nanoparticles mPPF/PFOB have good biocompatibility and can specifical y bind to breast cancer MCF-7 cel s in vitro and enhance the imaging capability.
RESUMO
Objective To investigate the intracellular delivery of siRNAs through the applications of ultrasound targeted microbubbles destruction(UTMD)and biodegradable nanoparticles carriers.Methods Preparation of nanoparticles with and without RGD sequences,parameters optimization via L16(45)orthogonal design,control experiments in groups of optimization,RGD targeted nanoparticles,non-RGD nanoparticles and blank control, and determinations by inverted fluorescence microscope and flow cytometry were performed.Results The uptake and fluorescence intensity of PC-3 cells in group of RGD targeted nanoparticle was (93.49±1.37)% and 34.28±2.06 respectively,and that in group of optimization was (88.33±1.24)% and 30.59±3.93 respectively(P＞0.05).Whereas the uptake and fluorescence intensity of PC-3 cells in group of non-RGD nanoparticles was(71.24±2.80)% and 18.39±0.90 respectively,and that in group of optimization was (84.78±2.13)% and 27.18±0.91 respectively(P＜0.05).ConclusionsThe applications of UTMD with RGD targted nanoparticles cannot increase the intracellular delivery of siRNAs.