低强度超声结合5-盐酸氨基乙酰丙酸治疗鼠舌癌

This topic adopts a non-invasive sonochemical treatment, using low-intensity ultrasound which was independent research and development by the Institute of Condensed Matter Science and Technology, Harbin Institute of Technology, combined with 5-HCl-aminolevulinic acid (5-ALA) on rat tongue cancer model for short-and long-term treatment. Filter out the appropriate drug dose and organ metabolism using high performance liquid chromatography-fluorescence detection from the animal level, determine the destruction of cancer cells sensitive threshold and radiation time (dose-effect relationship, time-effect relationship), and clear the superiority. Through methods such as morphology, pathology, cell biology, biochemistry, revealing the variation of the transfer process of rat tongue cancer after treatment. Through the light and electron microscopy, observe the changes in morphology and structure of tissues and cells. Using immunofluorescence, in situ hybridization, RT-PCR and Western blot technology, clear the ways that tumor cell death and the possible mechanism from the cellular and molecular levels, as well as the internal regulating relations between the tumor microenvironment and tumor cells during the outcome. Provide a scientific basis for the establishment of the biological evaluation method of low-intensity ultrasound treatment for solid tumors.

本项目采用非侵入性的声化学治疗方法，应用由哈尔滨工业大学凝聚态科学与技术研究所自主研发的低强度超声联合5-盐酸氨基乙酰丙酸（5-ALA）对鼠舌癌模型进行短、长期治疗。利用高效液相色谱-荧光检测法，从动物整体水平筛选出超声与5-ALA浓度等参数，确定杀伤癌细胞的敏感阈值和触发位点（量-效关系、时-效关系），明确其优效性；应用形态学、病理学、细胞生物学、生物化学等手段，揭示治疗鼠舌癌组织转归的变化规律。通过光、电镜观察组织和细胞形态和结构的变化；采用免疫荧光法、原位杂交、RT-PCR和Westernblot等技术，从细胞和分子水平阐明促进肿瘤细胞死亡的方式及可能作用机制，以及转归过程中肿瘤微环境和肿瘤细胞之间的内在调节关系，为建立低强度超声治疗肿瘤疗效的生物学评价方法提供科学依据。

本项目筛选出对正常细胞系（293T cell和L6 cell）无毒性作用的超声阈值与 5-盐酸氨基乙酰丙酸（5-ALA） 浓度等参数，确定最优效性（量-效关系和时-效关系）。证实了低强度超声（LIUS）联合5-ALA（LIUS-ALA）在抗血管生成和肿瘤生长抑制方面有协同作用。作为一种替代疗法，LIUS-ALA在实施肿瘤杀伤与诱导抗肿瘤免疫应答方面具有优势。LIUS-ALA能够促进肿瘤细胞释放DAMP，诱导DCs的成熟及向引流淋巴结的募集，促使局部引流淋巴结（TDLN）的空间结构发生重塑，并且逆转了TDLN的免疫耐受状态，激活T细胞，产生大量的效应T细胞并迁移至肿瘤部位杀伤肿瘤细胞。结果表明，低强度超声联合5-ALA不仅能抑制肿瘤、调节改善转归过程中肿瘤微环境和肿瘤细胞之间的内在关系，还能最大限度地减轻对正常组织的损伤，增加了药物靶向作用。LIUS-ALA衍生的抗肿瘤免疫方面的研究可能是极有前途的研发治疗仪。