Celastrol and Apoptin Mutant Exert Synergistic Anti-tumor Effects by Enhancing Nur77-induced Apoptosis Pathway

doi:10.13560/j.cnki.biotech.bull.1985.2019-0944

Abstract

Abstract: Both celastrol and apoptin can exert anti-tumor effect through the Nur77 pathway. We named a truncated variant of apoptin as tApoptin. tApoptin co-administered with esculentoside A（EsA）is known simply as tApoptinE. Combination of cleastrol with tApoptinE is applied to enhance the inhibiting tumor cell growth activity of these two drugs,and the molecular mechanism for these two drugs synergistically exerting anti-tumor ability is studied. The cytotoxicity of the combination of celastrol with tApoptinE on tumor cells was detected by MTT assay,and the molecular mechanism of cell apoptosis was observed by flow cytometry and Western blot analysis. The results showed that celastrol whose concentration is above 300 nmol/L could effectively increase the anti-tumor activity of tApoptinE in cell line SMMC-7721 in comparison with the free tApoptinE. Under the combined conditions above,the combined index values of all combined groups were < 1,which showed that celastrol and tApoptinE synergistically inhibit the growth of tumor cells in vitro in this case. In addition,600 nmol/L celastrol in combination with tApoptinE had the best synergistic effect. Flow cytometry analysis revealed that the synergistic effect was due to cell apoptosis enhancement. Western blot analysis showed that after combined administration,celastrol and tApoptinE synergistically targeted their common Nur77 pathway which will have Nur77 phosphorylated. And the phosphorylated Nur77 can significantly regulate the expression of Caspase and Bcl-2 family proteins,hence enhancing the induction of apoptosis. Therefore,the combination of celastrol and tApoptinE can further induce tumor cell apoptosis by enhancing the apoptosis pathway induced by Nur77,so as to exert synergistic anti-tumor effect.

Key words: celastrol, apoptin, Nur77, synergistic anti-tumor, cell apoptosis

YIN Xiao-meng, CAO Xue-wei, WANG Fu-jun, ZHAO Jian, ZHANG Hui-zhan. Celastrol and Apoptin Mutant Exert Synergistic Anti-tumor Effects by Enhancing Nur77-induced Apoptosis Pathway[J]. Biotechnology Bulletin, 2020, 36(7): 119-129.

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