【作者】 王燕春；

【导师】 张国才；

【作者基本信息】 吉林大学 ， 预防兽医学， 2011， 硕士

【摘要】 为了深入研究旋毛虫抗肿瘤的的作用,本试验先在体外利用MTT和细胞凋亡的检测的方法初步确定了旋毛虫可溶性抗原可以使Hela细胞发生凋亡,在用ELISA方法初步确定了旋毛虫与Hela细胞间存在共同抗原,并利用SDS-PAGE和Western-blotting方法测得了相关抗原的分子量。运用离心超滤装置及透析等技术对相关抗原进行了分离,FITC-免疫组化法对该抗原进行了定位分析。在此基础上,建立了Hela细胞裸鼠移植瘤动物模型,并对旋毛虫与Hela细胞相关抗原抗体进行了抗肿瘤实验。实验结果显示:通过MTT法和细胞凋亡实验证明,旋毛虫可溶性抗原可以对Hela细胞的增殖具有抑制作用,ELISA法初步证明旋毛虫与Hela细胞间存在较强的免疫交叉反应;利用SDS-PAGE和Western-blotting方法获得了旋毛虫抗原与Hela细胞相关抗原的分子量分别是40KDa和55KDa。免疫组化定位分析该相关抗原位于Hela细胞表面抗原和旋毛虫的消化道上皮细胞膜上;应用相关抗原抗体在裸鼠体内进行抗肿瘤试验表明,相关抗原抗体具有抗肿瘤作用,抑瘤率为82.3%,且效果略好于可溶性抗原抗体组。首次以旋毛虫与Hela细胞相关抗原抗体为切入点研究旋毛虫抗肿瘤的机理,为旋毛虫抗肿瘤机制的研究与开发提供了理论依据。更多还原

【Abstract】 Trichinosis is a globalized zoonotic parasitic disease.Trichinella spiralis that is the most frequent and food-transmitted helminth zoonosis can infect the human and 150 kinds of animals,what is more,cause people’s death.In the previous study ,some scholars discovered that when the hosts are infected with Trichinella spiralis their immunity can be enhanced,however,when hosts are infected with other nematodes,the host’s immunity is not remarkblely improved. It is suggested that Trichinella spiralis can activate the host’s immune system,moreover,it can contain some antitumor components.In order to study antitumor mechanism of Trichinella spiralis,the experiments were designed to investigate the anti-tumor mechanism of Trichinella spiralis and Hela cell related antigens-antibodies,furthermore,pave the way for inventing some new anti-tumor biological agent.MTT assay and cell apoptosis experiments showed that the soluble antigen of Trichinella spiralis on the proliferation of Hela cells inhibit the antigen concentration was 177μg/μl, apoptosis rate was 48.2%, inhibition was the best.ELISA method proved soluble antigen of Trichinella spiralis and Hela cells produce soluble antigen antibody cross-reactivity, Hela cells with soluble antigen soluble antigen of Trichinella antibodies produced by cross-reaction, initially identified Trichinella Hela cells with immune cross-reaction between the Common antigen.SDS-PAGE electrophoresis showed that the soluble antigen of Trichinella and Hela cells the same size as the soluble antigen is present antigen are 83KDa, 55KDa, 40KDa, 28.5KDa, 20KDa, 17.6KDa and 14.4KDa. Western-blot showed that protein in the immune response occurs, the size of 40 KDa. Hela cells at exponential phase of growth were collected and spreaded uniformly on POLY-L-LYSINE treated glass slide. Experimental anti-tumor-associated antigen research: experiments on tumor-bearing mice to observe the anti-tumor effects.The related antigens could be the basis to study the anti-tumor mechanism of Trichinella spiralis.The results show that Trichinella -associated antigen is located on the digestive tract and the Hell cell surface. The model of Hela cells in nude mice is successfully established. Trichinella antibodies and soluble antigen Hela cell-associated antigen antibody can reduce tumor xenografts in nude mice, tumor inhibition rates were 52.2% and 82.3%.更多还原