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Biological effects and mechanism of adenanthin on multiple myeloma cells

XIAO Xin-hua1, YU Miao1, WU Yun-zhao2, WU Ying-li2, LIU Wei1   

  1. 1. Pathology Center, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China; 2. Department of Pathophysiology, The Key Laboratory of Cell Differentiation and Apoptosis of the Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2017-04-28 Published:2017-05-04
  • Supported by:

    National Natural Science Foundation of China, 81570118; Foundation of Science and Technology Committee of Shanghai Municipality, 15401901800;
    Shanghai Municipal Commission of Health and Family Planning,201540226; National University Student Innovation Program of Shanghai Jiao Tong University


Objective · To explore the biologic effect and mechanism of adenanthin (Aden) on multiple myeloma (MM) cells. Methods · MM cells, H929 and U266 were treated with various dose of Aden for different time, and the density and viability of MM cells were detected by trypan blue exclusion assay. After H929 and U266 cells were treated with various dose of Aden for 24 hours, cell growth inhibition was examined by CCK8 assay, and cell apoptosis was examined by AnnexinV-APC/PI staining assay. Apoptosis related proteins, NF-κB signaling pathway associated proteins and the NFκB regulated proteins were detected by Western blotting. The effect of Aden on the thermal stability of IKKβ protein was determined by CETSA assay. Results · Trypan blue exclusion results showed that Aden inhibited cell growth and reduced cell viability in concentration and time dependent manners. U266 was more sensitive than H929 when exposed to the same concentration of Aden. The CCK8 results showed that Aden inhibited the growth of H929 and U266 cells in a concentration dependent manner. Flow cytometry results suggested that Aden induced a low apoptosis rate of MM cells. Moreover, cleavage of caspase3 and PARP were detected in U266 cells but not in H929 cells. CETSA assay indicated that Aden decreased the thermal stability of IKKβ. Expression of p-p65 and p-IκBα proteins decreased in MM cells treated with Aden. Conclusion · Aden significantly inhibits MM cell proliferation by inhibiting NF-κB activation through interacting with IKKβ. Aden has little effect on apoptosis of MM cells.

Key words: multiple myeloma, adenanthin, proliferation inhibition, cell apoptosis, NF-κB signaling pathway