非致病性大肠杆菌配合溶组织内阿米巴使肝癌衍变成肝脓肿的互补效应及机制

The appropriate animal models of liver cancer will be set up. Entamoeba histolytica trophozoites and E. coli will be inoculated in the liver cancer tissue. Complementary Effect and mechanism of non-pathogenic E. coli in coordination with entamoeba histolytica to change liver tumor into liver abscess will be investigated. This will include the conversion rate of hepatic abscess form liver cancer，situation of the treatment,survival, pathological of tissues and cells, Immune-related molecular biology research and security of treament, etc. Our working hypothesis on the possible mechanisms includes: 1. E. coli may provide food and improve the living environment for Entamoeba histolytica , promote their breeding, and enhance their aggregation, swallow , soluble tumor activity. 2. E. coli and Entamoeba synergies stimulate immune function to a "bystander effect" inside the tumor environment changes more quickly and more conducive to tumor cell growth;3. In terms of security, there were less chances of causing bacteremia and distant metastasis because of that Entamoeba histolytica and E. coli are facultative anaerobic organisms which much more easy to accumulate in the tumor. We will open a new liver cancer biological treatment window to establish a peaceful evolution of a malignant solid tumors converting to microbial infection abscess. Through this research project ,we would like to made the no cure disease into a treatable disease,to promote the development of the biological treatment of liver cancer, in order to achieve new breakthroughs in the field of translational medicine.

本研究拟建立适宜的肝癌动物模型，将溶组织内阿米巴滋养体与非致病性大肠杆菌同时接种于肝癌组织中，从动物肝癌的肝脓疡转化率；治疗效果与生存情况；组织细胞病理形态；免疫相关分子生物学及安全性等方面研究大肠杆菌配合溶组织内阿米巴使肝癌衍化成肝脓肿的互补效应及机制。假设的作用机制包括：1、大肠杆菌可为溶组织内阿米巴提供食物及改善生存环境，促进其繁殖，并使其凝集、吞噬、溶解肿瘤的活力明显增强；2、大肠杆菌与溶组织内阿米巴的协同作用，刺激机体免疫功能产生"旁观效应"使肿瘤内部环境改变更加迅速更加不利于肿瘤细胞生长；3、在安全性方面，大肠杆菌与溶组织内阿米巴均为兼性厌氧生物，易于蓄积在瘤内，造成菌血症及远处转移的机会较少。通过项目研究，建立一种将恶性实体瘤转化为微生物感染性脓肿的和平演变的肝癌生物治疗模式，将不可治性疾病转化为可治性疾病，促进肝癌的生物学治疗的发展，为转化医学领域实现新的突破。

目的：研究溶组织内阿米巴诱导肝细胞肝癌自然良性回归为肝脓肿的作用及机制，探索肝癌治疗的新模式。.内容：1.溶组织内阿米巴体外抗肿瘤作用及机制。. 2.建立人肝癌细胞HepG2裸鼠种植性肝癌模型。. 3.溶组织内阿米巴体内抗肿瘤的作用及机制。.结果：与溶组织内阿米巴滋养体共培养的肝癌细胞在30小时后基本消失；肝癌细胞克隆形成受到明显抑制，流式细胞检测发现肝癌细胞凋亡率为80%，Western Blot实验结果发现溶组织内阿米巴滋养体可促进肝癌细胞的凋亡并上调死亡受体FASL的表达；动物实验表明：阿米巴干预组，溶组织内阿米巴能使种植性肝癌不同程度的转化为阿米巴肝脓肿(9/10)，对照组未发现肝脓肿（0/11）,病理切片HE染色发现有滋养体，残存的癌组织明显减少，与对照组比较有显著性差异(P<0.05)。肿块脓液涂片镜下观察有活动溶组织内阿米巴滋养体。脾脏、肾脏、心脏、肺、脑的组织形态学无变化，生理盐水对照组肝肿块为肝癌组织。阿米巴干预组血培养没有发现细菌生长。.结论：1、溶组织内阿米巴体外能明显抑制肝癌细胞的增殖；2、溶组织内阿米巴具有诱导肝癌细胞凋亡的作用，其机制与上调死亡受体FASL的表达有关；3、野生型溶组织内阿米巴在大肠杆菌的协同下，瘤内接种，能使裸鼠人肝癌细胞种植性肝癌转化为阿米巴肝脓肿，对其它组织器官没有发现侵害。

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