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Juglanthraquinone C, a novel natural compound derived from Juglans mandshurica Maxim, induces S phase arrest and apoptosis in HepG2 cells

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Abstract

Juglanthraquinone C (1,5-dihydroxy-9,10-anthraquinone-3-carboxylic acid, JC), a naturally occurring anthraquinone isolated from the stem bark of Juglans mandshurica, shows strong cytotoxicity in various human cancer cells in vitro. Here, we first performed a structure–activity relationship study of six anthraquinone compounds (JC, rhein, emodin, aloe-emodin, physcion and chrysophanol) to exploit the relationship between their structural features and activity. The results showed that JC exhibited the strongest cytotoxicity of all compounds evaluated. Next, we used JC to treat several human cancer cell lines and found that JC showed an inhibitory effect on cell viability in dose-dependent (2.5–10 μg/ml JC) and time-dependent (24–48 h) manners. Importantly, the inhibitory effect of JC on HepG2 (human hepatocellular carcinoma) cells was more significant as shown by an IC50 value of 9 ± 1.4 μg/ml, and 36 ± 1.2 μg/ml in L02 (human normal liver) cells. Further study suggested that JC-induced inhibition HepG2 cell proliferation was associated with S phase arrest, decreased protein expression of proliferation marker Ki67, cyclin A and cyclin-dependent kinase (CDK) 2, and increased expression of cyclin E and CDK inhibitory protein Cip1/p21. In addition, JC significantly triggered apoptosis in HepG2 cells, which was characterized by increased chromatin condensation and DNA fragmentation, activation of caspase-9 and -3, and induction of a higher Bax/Bcl2 ratio. Collectively, our study demonstrated that JC can efficiently inhibit proliferation and induce apoptosis in HepG2 cells.

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Abbreviations

DAPI:

4,6-Diamidino-2-phenylindole

DMSO:

Dimethyl sulfoxide

DMEM:

Dulbecco’s modified Eagle’s medium

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

FBS:

Fetal bovine serum

JC:

Juglanthraquinone C

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide

PBS:

Phosphate balanced solution

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

PFA:

Paraformaldehyde

RT:

Room temperature

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (Grant Nos. 31170324 and 31070318), The Fundamental Research Funds for the Central Universities, and the Research Foundation of Jilin Provincial Science & Technology Development (Grant Nos. 20100911, 20110938, 201201065 and 20110711), The Science Foundation for Young Teachers of Northeast Normal University (C03020705).

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Authors and Affiliations

  1. National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun, 130024, China

    Yao Yao, Yu-Wei Zhang, Yong-Li Bao, Yu Zhang, Chun-Lei Yu, Yin Wu & Guan-Nan Wang

  2. Department of Hand Surgery, China–Japan Union Hospital, Jilin University, Changchun, 130031, China

    Biao Liu

  3. Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University, Changchun, 130024, China

    Lu-Guo Sun, Hua Lin, Li-Hua Zheng, Ying Sun & Yu-Xin Li

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  1. Yao Yao
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  2. Yu-Wei Zhang
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Correspondence to Biao Liu, Yong-Li Bao or Yu-Xin Li.

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Yao, Y., Zhang, YW., Sun, LG. et al. Juglanthraquinone C, a novel natural compound derived from Juglans mandshurica Maxim, induces S phase arrest and apoptosis in HepG2 cells. Apoptosis 17, 832–841 (2012). https://doi.org/10.1007/s10495-012-0722-5

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  • DOI: https://doi.org/10.1007/s10495-012-0722-5

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