The efficient production of ganglioside analogues was accomplished using RERF-LC-AI cells cultured in HYPERFlask (High Yield PERformance Flask). Eight kinds of ganglioside analogues (GM3, GM2, sialylparagloboside, GD3, di-sialylated lacto-N-tetraose, and another three kinds of analogues with intricate structures) were synthesized by the saccharide primer method using lung squamous-cell carcinoma line RERF-LC-AI and 12-azidododecyl β-lactoside primer. The yield for each analogue obtained using HYPERFlask was higher than yields obtained from 100-mm dishes.

No abstract

Introduction: Photodynamic therapy (PDT) has already been established as anti-cancer therapy. However, since some types of cancer proliferate multifocally and/or diffusely, topical irradiation of light by a conventional optical fiber results in an insufficient PDT effect. Moreover, using a conventional fiber, we cannot irradiate minimal tumors that escape from detection by conventional diagnostic approaches. When PDT is applied to the lesions of hollow organs, if we homogenously irradiate a wide area of the organ wall, undetectable minimal tumors and multifocally proliferating tumors can be treated simultaneously. In order to realize this concept, we have developed a novel fiber probe that provides homogenous light irradiation on the mucosa of hollow organs. In this study, we fabricated a homogenous irradiation fiber probe (HIFP) for illuminating the bladder by the use of state-of-the-art processing technology (torsional deformation of a fiber core, conical shaping of fiber tip, etc). We evaluated the PDT effect on a rat orthotopic bladder tumor model using the HIFP and a dendrimer phthalocyanine micelle (DPc/m) that we developed as a DDS-type photosensitizer. Materials and Methods: The property of spatial distribution of light emitted from the HIFP was measured. Using the HIFP, PDT was carried out on a rat orthotopic bladder tumor model 24 h after administration of a photosensitizer (Photofrin or DPc/M) and PDT effects were pathologically evaluated. Results: Preliminarily examination of ultrasonic imaging revealed that the shape of the rat bladder approximates an ellipsoidal body with a long radius of 7.5 mm and a short radius of 5 mm. Analysis of spatial distribution showed that the light emitted from the HIFP covered 90% of the whole luminal area of the approximate ellipsoidal body, while the light emitted from a control fiber (CF) (NA=0.41; flat cleaved end) covered only 15%. Pathological study of the PDT-treated rat using the CF revealed that an anti-tumor effect occurred only in the local area proximate to the focal point of the CF with no anti-tumor effect in tumors in other areas of the bladder wall. In contrast, pathological study of the PDT-treated rat using the HIFP revealed that light was distributed over 60 −80% of the mucosa of the bladder and tumor proliferation was widely suppressed. Conclusion: Widespread and homogenous irradiation in the mucosa of the bladder was achieved using the newly developed fiber probe. PDT using a combination of the fiber probe and DPc/M enhanced the anti-tumor effect without bladder shrinking. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 501.

，胡 尉之

[Background and purpose] Direct observation of pathological lesions by a relatively noninvasive approach is important for understanding pathophysiological mechanism and for assessing the objective response to therapeutic interventions. On the other hand, it is very difficult to perform direct and relatively noninvasive observation of minimal lesions inside a diseased model using a small live animal. However, recent technological developments in imaging, especially significant progress in fluorescent methods and in endoscopic examinations, have enabled detection and tracing of micro-lesions inside small animal models. We have developed a fluorescence imaging technique using high-resolution ultrathin endoscope, which is accessible to hollow organs of small animals. In addition, we have established an orthotopic rat bladder cancer model in which the cancer cells express a fluorescent protein. The aim of this study was to establish a high-resolution imaging method for visualizing the vascular distribution and mucosal morphology of a bladder tumor. [Materials and Methods] GFP-expressing bladder tumor in a rat orthotopic model: Firstly, we established a cell line with stable integration of GFP constructs that were produced by transfection of pTurboGFP into rat bladder cells (AY27). The stable AY27+tGFP populations were purified by repeated selections using Geneticin (G418). AY27-tGFP cells were implanted in the bladders of female Fischer F344 rats, resulting in development of submillimeter-sized bladder tumors. Development of fluorescence ultrathin endoscopy: We developed a fluorescence ultrathin endoscopic system installed with a 180 W Xenon lamp with excitation filters and 3CCD camera with emission filters. The outer diameter of the ultrathin endoscopic fiber is only 0.8 mm, but its number of pixels reaches 15,000, resulting in a spatial resolution of 50μm. [Results] Fluorescence from AY27-tGFP cells implanted in the bladder was clearly visualized by the ultrathin endoscope placed in the bladder by the transurethral approach. Furthermore, when fluorescence-probed tumor-directive micelles were injected (i.v.) into the animals as a model of drug delivery, fluorescence from the probe agents was visualized in accordance with tumor distribution. In both cases, we detected fluorescence from tumors of less than 1 mm in size. [Conclusion] We established an orthotopic GFP-tumor rat model using a GFP-expressing tumor cell line (AY27-tGFP). Submillimeter tumors in this model were visualized by the newly developed ultrathin endoscopic system. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5235.