Mutant and wild-type TNF.doc

 PAGE \* MERGEFORMAT 8 Mutant and wild-type TNF [Keywords:], tumor Apoptosis of tumor cells induced by mutant and wild type TNFα [Abstract] AIM: To compare the apoptosisinduced ability of recombinant human mutant type 471 rhTNFα (mt 471rhTNFα ) with that of wild type rhTNFα (wt rhTNFα ) and to study the apoptotic mechanism induced by mt 471rhTNFα . METHODS: The apoptosis of ZR751 cells, a breast cancer cell line, induced by mt 471rhTNFα or wt rhTNFα was analyzed and compared by 20 g / L agarose gel electrophoresis and flow cytometry techniques. The activation profiles of nuclear factor NFκ B in ZR751 cells untreated and treated by mt 471rhTNFα or wt rhTNFα were detected using Trans AMTM NFκ B p65 kit based on ELISA for further study of the apoptotic mechanism induced by mt 471rhTNFα . RESULTS: The results from 20 g / L agarose gel electrophoresis of genomic DNA indicated that ZR751 cells treated by mt 471rhTNFα provided a typical apoptotic ladder pattern of DNA fragmentation, whereas weakened ladders were observed in ZR751 cells treated by wt rhTNFα . Flow cytometry showed that the cell apoptotic rate in mt 471rhTNFα treated group was 43%, but 25% in wt rhTNFα treated group. The mt 471rhTNFα held a strong apoptosisinducible ability. The results of DNAbinding activity of NFκ B showed that NFκ B activation induced by wt rhTNFα was significantly higher than that of mt 471rhTNFα when the protein concentration of mt 471rhTNFα or wt rhTNFα reached 50 μ g / L (P = 0.002). CONCLUSION: The apoptosisinduced ability of mt 471rhTNFα is superior to that of wt rhTNFα . One of the possible reasons for the enhanced apoptosisinduced ability may be that NFκ B activation is inhibited in tumor cells treated with mt 471rhTNFα . [Keywords] neoplasms; mutant type 471rhTNFα ; wild type rhTNFα ; NFkappaB; apoptosis [Abstract] Objective: To compare recombinant human mutant 471rhTNFα (mt 471rhTNFα ) with wild-type rhTNFα (wt rhTNFα )-induced apoptosis of tumor cell