Theranostics 2019; 9(26):8109-8126. doi:10.7150/thno.35378

Research Paper

Hepatomas are exquisitely sensitive to pharmacologic ascorbate (P-AscH-)

Xuan Zhang1#, Tiefu Liu1#, Zehuan Li2#, Yanling Feng1, Christopher Corpe3, Shanshan Liu1, Jingpu Zhang1, Xiaomeng He1, Feng Liu1, Li Xu1, Longqiang Shen1, Shun Li1, Qianlin Xia4, Xiuhua Peng1, Xiaohui Zhou1, Weiping Chen5, Xiaoyan Zhang1, Jianqing Xu1, Jin Wang1✉

1. Shanghai Public Health Clinical Center, Fudan University, 2901 Caolang Road, Jinshan District, Shanghai 201508, China;
2. Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, China;
3. King's College London, London, Nutritional Science Department, 150 Stamford street, waterloo, London, SE19NH, United Kingdom;
4. Department of Laboratory Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China;
5. Genomics Core, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
# These authors contributed equally to this work

This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions.
Citation:
Zhang X, Liu T, Li Z, Feng Y, Corpe C, Liu S, Zhang J, He X, Liu F, Xu L, Shen L, Li S, Xia Q, Peng X, Zhou X, Chen W, Zhang X, Xu J, Wang J. Hepatomas are exquisitely sensitive to pharmacologic ascorbate (P-AscH-). Theranostics 2019; 9(26):8109-8126. doi:10.7150/thno.35378. Available from http://www.thno.org/v09p8109.htm

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Abstract

Rationale: Ascorbate is an essential micronutrient known for redox functions at normal physiologic concentrations. In recent decades, pharmacological ascorbate has been found to selectively kill tumour cells. However, the dosing frequency of pharmacologic ascorbate in humans has not yet been defined.

Methods: We determined that among five hepatic cell lines, Huh-7 cells were the most sensitive to ascorbate. The effects of high-dose ascorbate on hepatoma were therefore assessed using Huh-7 cells and xenograft tumour mouse model.

Results: In Huh-7 cells, ascorbate induced a significant increase in the percentage of cells in the G0/G1 phase, apoptosis and intracellular levels of ROS. High doses of ascorbate (4.0 pmol cell-1), but not low doses of ascorbate (1.0 pmol cell-1), also served as a pro-drug that killed hepatoma cells by altering mitochondrial respiration. Furthermore, in a Huh-7 cell xenograft tumour mouse model, intraperitoneal injection of ascorbate (4.0 g/kg/3 days) but not a lower dose of ascorbate (2.0 g/kg/3 days) significantly inhibited tumour growth. Gene array analysis of HCC tumour tissue from xenograft mice given IP ascorbate (4.0 g/kg/3 days) identified changes in the transcript levels of 192 genes/ncRNAs involved in insulin receptor signalling, metabolism and mitochondrial respiration. Consistent with the array data, gene expression levels of AGER, DGKK, ASB2, TCP10L2, Lnc-ALCAM-3, and Lnc-TGFBR2-1 were increased 2.05-11.35 fold in HCC tumour tissue samples from mice treated with high-dose ascorbate, and IHC staining analysis also verified that AGER/RAGE and DGKK proteins were up-regulated, which implied that AGER/RAGE and DGKK activation might be related to oxidative stress, leading to hepatoma cell death.

Conclusions: Our studies identified multiple mechanisms are responsible for the anti-tumour activity of ascorbate and suggest high doses of ascorbate with less frequency will act as a novel therapeutic agent for liver cancer in vivo.

Keywords: Ascorbate, vitamin C, cancer, hepatoma, ROS, mitochondrial respiration.