20(S)-25-methoxyl-dammarane-3β, 12β, 20-triol negatively regulates activation of STAT3 and ERK pathways and exhibits anti-cancer effects in HepG2 cells
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The pro-inflammatory cytokine interleukin 6 (IL-6), via activating its downstream JAK/STAT3 and Ras/ERK signaling pathways, is involved in cell growth, proliferation and anti-apoptotic activities in various malignancies. To screen inhibitors of IL-6 signaling, we constructed a STAT3 and ERK dual-pathway responsive luciferase reporter vector (Co.RE). Among several candidates, the natural compound 20(S)-25-methoxyl-dammarane-3β, 12β, 20-triol (25-OCH3-PPD, GS25) was identified to clearly inhibit the luciferase activity of Co.RE. GS25 was confirmed to indeed inhibit activation of both STAT3 and ERK pathways and expression of downstream target genes of IL-6, and to predominantly decrease the viability of HepG2 cells via induction of cell cycle arrest and apoptosis. Interestingly, GS25 showed preferential inhibition of HepG2 cell viability relative to normal liver L02 cells. Further investigation showed that GS25 could not induce apoptosis and block activation of STAT3 and ERK pathways in L02 cells as efficiently as in HepG2 cells, which may result in differential effects of GS25 on malignant and normal liver cells. In addition, GS25 was found to potently suppress the expression of endogenous STAT3 at a higher concentration and dramatically induce p38 phosphorylation in HepG2 cells, which could mediate its anti-cancer effects. Finally, we demonstrated that GS25 also inhibited tumor growth in HepG2 xenograft mice. Taken together, these findings indicate that GS25 elicits its anti-cancer effects on HepG2 cells through multiple mechanisms and has the potential to be used as an inhibitor of IL-6 signaling. Thus, GS25 may be developed as a treatment for hepatocarcinoma with low toxicity on normal liver tissues as well as other inflammation-associated diseases.
KeywordsSTAT3 ERK 25-OCH3-PPD Apoptosis Anti-tumor effects HepG2
We would like to thank Xiu-Li Gao and Wen-Jing Zhang for helpful guidance in data analysis and experimental methods; Yi-Meng Wang, Xiao-Jia Liu and Xin Xu for animal experiments; Li-Hua Zheng, Ying Sun and Guan-Nan Wang for technical support. We also thank the supporting grants from the National Natural Science Foundation of China (No. 81372870), National Key New Drug Creation and Manufacturing Program of Ministry of Science and Technology (No. 2013ZX09103003004), Fundamental Research Funds for the Central Universities, Grant of Jilin Province Science & Technology Committee (Nos. 20110711, 20150204038YY, 20130206072YY, YYZX201121, 20130201008ZY, 20150309003YY and 20170414028GH), Grant of Administration of Traditional Chinese Medicine of Jilin Province (2014-ZDR) and Grant of Changchun Science & Technology Committee (Nos. 2014070, 2013314).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Animal experiments were approved by the Ethics Committee of our institution and were performed in accordance with Regulations for the Administration of Affairs Concerning Experimental Animals approved by the State Council and promulgated by Decree No. 2 of the State Science and Technology Commission (1988). All applicable international, national and/or institutional guidelines for the care and use of animals were followed.
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