Anti-tumor Effect On Hepa1-6 Hepatoma Carcinoma Cell Of Trichinella Spirialis

It was found that 6 350 000 new cases of malignancy in the world every year, and 260 000 cases were liver cancer .42.5% of whole world liver cancer cases were found in our country. was promulgated by Department of Health in Dec .2003 . This program defined that eight kinds of cancers (lung cancer, liver cancer, carcinoma of stomach, carcinoma of esophagus, colorectal cancer, cancer of the cervix, cancer of the cervix, nasopharyngeal carcinoma) are emphasis Cancer to prevent and treat, and above 80% cancer death were cause of those eight cancers. Liver cancer, called the king of cancer, made the highest death rate in all cancer.Along with the development of the research on tumorigenesis molecule mechanism and biotechnology, biotherapy is becoming the most potential treatment mode on combined therapy of tumour.It was found in some research that some viruses and parasites make some damage on organism, but at the same time they can do some benefit also. For instance Newcastle disease virus, adeno-associated virus, Oncolytic virus, trichina, Amoeba trophozoite, and plasmodium do have some function on Anti-tumor. Trichinos is a globalized zoontic parasitic disease. Trichinella spiralis is the most frequent and food-transmitted helminth zoonosis. In the previous study , some scholars discovered that when the hosts are infested with Trichinella spiralis their immunity can be enhanced, but other nematodes have no such function. It is suggested that Trichinella spiralis can activate the host's immune system , moreover, and there are some antitumor components in it. Zhang Jingzhi et al found that the fos-related antigens between SP2/0 myeloma cell and Trichinella spiralis is Tropomyosin, which includes 284 amino acid sequences of fos-related antigens. At least three pairs of similar molecular weight protein zones were found in both A549 lung cancer cell antigen and Trichinella spiralis antigen by Gong Pengtao et al. The molecular weights of the three pairs of protein zones are 97.4kDa, 68kDa and 46kDa separately.In order to found whether the fos-related antigen exist between Trichinella spiralis and Hepa1-6 hepatoma carcinoma cell, this reserch includes four following sections: Observing the effect of Trichinella spiralis polypide and Trichinella spiralis antigen on Hepa1-6 hepatoma carcinoma cell growth to find if there is a depressant effect of Trichinella spiralis polypide and Trichinella spiralis antigen on Hepa1-6. Using enzyme linked immunosorbent assay (ELISA) to found if the fos-related antigen exist between Trichinella spiralis and Hepa1-6 hepatoma carcinoma cell, Determining the molecular weight of fos-related antigen by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.To study the anti-tumor effect of Trichinella spiralis on Hepa1-6 hepatoma carcinoma cell in mice, we transplant Hepa1-6 hepatoma carcinoma cell to C57BL/6 mice then vaccinate C57BL/6 mice with weakened(weakened by UV ray and Co60) or normal Trichinella spiralis 7 d later in the first group, vaccinate C57BL/6 mice with weakened or normal Trichinella spiralis then transplant Hepa1-6 hepatoma carcinoma cell to C57BL/6 mice 11 d later in the second group, and mice with transplant Hepa1-6 hepatoma carcinoma cell only is control group. Then we measured volume and weight of the tumor, analysised inhibitory rate, and determined immunological index. We found that Trichinella spiralis has inhibitory effect on the growth of Hepa1-6 hepatoma carcinoma cell in mice .Comparing with the control group, the tumor volume and weight in normal Trichinella spiralis vaccinated, ultraviolet weakened Trichinella spiralis vaccinated and Co60 weakened Trichinella spiralis vaccinated mice were significantly decreased. And the inhibitory effect of normal Trichinella spiralis vaccinated is the best. The higher ratios of CD4+ to CD8+ in three vaccinated groups indicated that Trichinella spiralis can stimulate the immunity function, strengthen the ability in anti-tumor effect.To study the inhibitory action on Hepa1-6 hepatoma carcinoma cell of different Trichinella spiralis antigenes(Trichinella spiralis soluble crude antigen and ES antigen ), C57BL/6 mice were vaccinated with different Trichinella spiralis antigen and then transplanted with Hepa1-6 hepatoma carcinoma cell . After that we measured volume and weight of the tumorous, analysised inhibitory rate, determined immunological index. We found that the tumor volume and weight in two vaccinated groups were obviously smaller than control group, but there is no remarkably difference between the two vaccinated groups ; Comparing with control group, the numerus of CD3+,CD4+ and CD4+/CD8+ of two vaccinated groups is significant increased. The results indicate that Trichinella spiralis soluble crude antigen and ES antigen both have the anti-tumor effect on Hepa1-6 hepatoma carcinoma cell.ELISA was used to determined whether Hepa1-6 hepatoma carcinoma antigen has cross-reaction with serum immunized with Trichinella spiralis and Trichinella spiralis antigen has cross-reaction with serum immunized with Hepa1-6 hepatoma carcinoma cell. The operation of ELISA was established according to basic procedure. We found that Trichinella spiralis larva in muscle and hepatoma carcinoma cellhave fos-related antigen that has cross-reaction. That was beneficial to determine the molecular weight of fos-related antigen.The components of hepatoma carcinoma cellantigen and Trichinella spiralis antigens were separated by SDS-PAGE. Western-blotting was used to further determine the molecular weight of fos-related antigen.The result indicate that fos-related antigen exist between hepatoma carcinoma cellantigen and Trichinella spiralis antigens, and fos-related antigen molecular weight have big different.