Inhibition Effect Of¹³¹I-CD133amb Association DDP On Liver In Vitro And Vivo

Objective:CD133mAb can connection CD133antigen in humanliver cancer. Study the inhibition effect of131I-CD133mAb on liver cancerwith β-ray which missive by131I.Part1Detecting of CD133+liver cancer cells in different human livercancer cells and tumor formation in nude miceMethods: Flow cytometry (FCM) was used to detect the expressionof CD133in Huh-7cell and HepG2cell. Immunohistochemistry was usedto detect the expression of CD133in tumors. CD133+and CD133-cellswere isolated from Huh-7cell and HepG2cell through magnetic-activatedcell sorting(MACS).CD133+expression in both sorted and unsorted cellswas analyzed by flow cytometry(FCM).Huh-7cells was cultural in CSCsculture-medium and CD133+expression was analyzed by flow cytometry.CD133+and CD133-cells were used to planted in nude mice.Result:The CD133expression in Huh-7was79.53%patency higherthan HepG2cells(4.33%). The result of Immunohistochemistry was that there are amount of brown yellow granules in Huh-7cell tumors, andHepG2cell tumors has a little, It was consistent with the result in vitro.The result of CD133+expression in sorted and unsorted Huh-7cells was99.28%and74.98%, which in HepG2cells was95.04%and3.73%. Theresult of CD133+expression in suspension culture Huh-7cells was96.39%.Several thousands CD133+cells can lead to tumorigenesis, but tenmillion CD133-cells can not lead to tumorigenesis.Conclusion: the expression of CD133in Huh-7cells was higherthan HepG2cells in vivo and in vitro. CD133+cells has the biologicalcharacteristics of tumor stem cells.Part2Inhibition effect of131I-CD133mAb association DDP on livercancer in vitro and in vivo.Methods: The monoclonal antibody CD133labeled with131I using thechloramines-T method and evaluate the stability. There were four groupsincluding131I-CD133mAb association DDP group, the131I-CD133mAbgroup, the DDP group and the blank group. The inhibitory effects ofdifferent treatments on the proliferation of human liver cancer cell lineHuh-7were measured by MTT assay and calculate each group,s IC50.Theanimal model was established by subcutaneous inoculation of human livercancer cell line Huh-7cells(5×106) in right front legs of BALA/c mice. Themodel mice were random allocation into four groups, all groups were treatmented by drugs every two days time for two weeks. The tumor sizeand volume were measured twice a week after treatment. After two weeksthe mice were sacrificed for pathological examination of the tumor. Theinhibitory rate was calculated and tumor apoptosis was observed by HEstaining.Result: The labeling ratio of the131I-CD133mAb was90.25%，theradiochemical purity was97.78%. The tumor inhibitory rate of131I-CD133mAb association DDP group was higher than other groups invivo. A great deal of tissue necrosis after treatment by131I-CD133mAbassociation DDP in HE staining. CD133+cells were specific binding by131I-CD133mAb and treaded by β-ray.Conclusion: The labeling ratio of the131I-CD133mAb was higher and131I-CD133mAb can containment the liver cancer in high expressionCD133antigen in vivo and in vitro.