Mechanism Research of Niclosamide Induced Maladjustment of BCAA through Circadian Disorder by Down-Regulating CLOCK
CHEN Tongzuan1#, DAI Ruixia2#, JIN Gaowei2, LIANG Qi2, ZHU Xiaohui2, DAI Jichen2, HONG Peibin2, ZHANG Jiaqi2, CHEN Tongke3, FANG Hezhi4, CHEN Xiangjian1, XIE Bintao5 *
Circadian rhythm is deemed a cyclical phenomenon of lives, while metabolic reprogramming exists in cancer cells at large. Some researches have found relationship between circadian and metabolic regulation. Niclosamide, a traditional anti-worm drugs, armed with properties to uncouple oxidative phosphorylation, is marching to the researching field of anti-tumour drugs. To illuminate mechanisms of its anti-tumour properties, cell counting, reactive oxygen species and apoptosis assay were performed to find that niclosamide could inhibit proliferation and promote apoptosis. Clock, Per, Dbp were selected as circadian differential expression genes through bioinformatics analyses, and expressions of these proteins were in disorder by Western blot, especially CLOCK, after treatment with Niclosamide. The study went deeper into discussing relevance between circadian disorder and metabolism through seahorse analyses and metabonomics analyses. Intracellular oxidative phosphorylation was tested to be suppressed after treatment with niclosamide in a concentration-dependent way. Besides, the importance of fumaric acid in mechanisms of niclosamide’s anti-tumour effect were also found. Western blot was performed to figure out that BCAA (branched amino acid)-related protein were inhibited after treatment with niclosamide, and combination of niclosamide with knockdown of CLOCK could further suppress the expression of BCAT1 (branched chain amino acid transaminase 1). Taken altogether, this research displayed that niclosamide could contribute to maladjustment of BCAA through circadian disorder to suppress tumour, which had a promising future as a new star of the anti-tumour.
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