Physcion, a naturally occurring anthraquinone derivative, induces apoptosis and autophagy in human nasopharyngeal carcinoma.
OBJECTIVE
Physcion is a major bioactive ingredient in the traditional Chinese medicine Radix et Rhizoma Rhei, which has an anthraquinone chemical structure and exhibits a variety of pharmacological activities including laxative, hepatoprotective, anti-inflammatory, anti-microbial and anti-proliferative effects. In this study we investigated the effect of physcion on human nasopharyngeal carcinoma in vitro and in vivo, as well as the mechanisms underlying the anti-tumor action.


METHODS
The nasopharyngeal carcinoma cell line CNE2 was treated with physcion, and cell viability was detected using MTT and colony formation assays. Flow cytometry was used to assess the cell cycle arrest, mitochondrial membrane potential loss, apoptosis, autophagy and intracellular ROS generation. Apoptotic cell death was also confirmed by a TUNEL assay. The expression of target or marker molecules was determined using Western blotting. The activity of caspase-3, 8, and 9 was detected with an ELISA kit. A xenograft murine model was used to evaluate the in vivo anti-tumor action of physcion, the mice were administered physcion (10, 20 mg·kg-1·d-1, ip) for 30 d.


RESULTS
Treatment with physcion (5, 10, and 20 μmol/L) dose-dependently suppressed the cell viability and colony formation in CNE2 cells. Physcion (10 and 20 μmol/L) dose-dependently blocked cell cycle progression at G1phase and induced both caspase-dependent apoptosis and autophagy in CNE2 cells. Furthermore, physcion treatment induced excessive ROS generation in CNE2 cells, and subsequently disrupted the miR-27a/ZBTB10 axis, resulting in repression of the transcription factor Sp1 that was involved in physcion-induced apoptosis and autophagy. Moreover, physcion-induced autophagy acted as a pro-apoptotic factor, and possibly contributed to physcion-induced apoptosis. In the xenograft murine model, administration of physcion dose-dependently suppressed the tumor growth without affecting the body weight. Furthermore, the anti-tumor effects of physcion were correlated with downregulation of Sp1 and suppression of miR-27a in the tumor tissues.


CONCLUSIONS
Physcion induces apoptosis and autophagy in human nasopharyngeal carcinoma by targeting Sp1, which was mediated by ROS/miR-27a/ZBTB10 signaling. The results suggest that physcion is a promising candidate for the treatment of human nasopharyngeal carcinoma.
Department of Otolaryngology, Qingdao Municipal Hospital, Qingdao 266011, China.
Ming-Jie
MJ
Pang
Department of Otolaryngology, Qingdao Municipal Hospital, Qingdao 266011, China.
Zhun
Z
Yang
Department of Oncology, Qingdao Municipal Hospital, Qingdao 266011, China.
Xing-Lin
XL
Zhang
Department of Otolaryngology, Qingdao Municipal Hospital, Qingdao 266011, China.
Zhao-Fang
ZF
Liu
Department of Otolaryngology, Qingdao Municipal Hospital, Qingdao 266011, China.
Jun
J
Fan
Department of Dermatology, Qingdao Municipal Hospital, Qingdao 266011, China.
Hong-Ying
HY
Zhang
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D045744:Cell Line, Tumor; D004642:Emodin / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D064593:Heterografts; D006801:Humans; D051379:Mice; D009303:Nasopharyngeal Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D009368:Neoplasm Transplantation; D017382:Reactive Oxygen Species / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism
D000972:Antineoplastic Agents, Phytogenic; D017382:Reactive Oxygen Species; D016329:Sp1 Transcription Factor; C108209:SP1 protein, human; C008905:physcione; D004642:Emodin
Oncogene. 2012 Aug 30;31(35):3973-88
22158040
Ann Anat. 2010 Sep 20;192(5):275-83
20810260
J Biol Chem. 2001 Mar 9;276(10):7549-58
11114309
BMC Cancer. 2002 Dec 10;2:35
12475396
Eur J Pharmacol. 2015 Oct 5;764:124-33
26144377
Cell Res. 2012 Jan;22(1):43-61
21912435
J Pharm Pharmacol. 2010 Sep;62(9):1158-66
20796195
Acta Pharmacol Sin. 2016 Feb;37(2):264-75
26707141
PLoS One. 2015 Feb 25;10 (2):e0117375
25714809
PLoS One. 2012;7(9):e44810
23024765
Biochem Biophys Res Commun. 2004 Jan 9;313(2):397-403
14684175
PLoS One. 2013 Oct 18;8(10):e78958
24205354
Int J Cancer. 2009 Nov 1;125(9):2066-76
19588484
J Oral Pathol Med. 2015 Nov;44(10):785-91
25475012
Nat Rev Mol Cell Biol. 2014 Feb;15(2):81-94
24401948
Autophagy. 2010 Nov;6(8):1214-5
20935512
PLoS One. 2012;7(7):e41779
22848600
Apoptosis. 2014 Aug;19(8):1177-89
24850099
J Surg Oncol. 1986 Nov;33(3):194-7
3773537
J Transl Med. 2014 Aug 07;12:222
25099028
Food Chem Toxicol. 2011 Feb;49(2):527-32
21134410
Cell. 1983 Mar;32(3):669-80
6187469
Microbiol Res. 2014 Apr;169(4):255-61
24071573
Chin Med J (Engl). 2015 Apr 5;128(7):941-7
25836616
Cell Death Dis. 2015 Jan 22;6:e1604
25611379
Blood. 2002 Nov 1;100(9):3325-32
12384434
Clin Otolaryngol Allied Sci. 2001 Apr;26(2):82-92
11309046
Int J Cancer. 2006 Sep 1;119(5):1125-35
16570274
Nat Rev Cancer. 2004 Oct;4(10):757-68
15510157
Free Radic Biol Med. 2015 Dec;89:452-65
26454086
J Cell Mol Med. 2015 Mar;19(3):630-41
25598321
J Cell Biol. 2005 Nov 21;171(4):603-14
16286508
BMC Cancer. 2014 Sep 18;14:678
25239093
Genes Dev. 2003 Oct 15;17(20):2481-95
14561771
J Cancer Prev. 2014 Dec;19(4):273-8
25574462
Nat Cell Biol. 2015 Nov;17(11):1484-96
26479318
Gene. 1999 Oct 1;238(2):291-300
10570957
J Biol Chem. 2002 Aug 9;277(32):28815-22
12052832
BMC Cancer. 2012 Nov 30;12:564
23194063
J Ethnopharmacol. 2009 Jul 30;124(3):571-5
19464353
Apoptosis. 2010 Jul;15(7):822-33
20349136
Planta Med. 2005 Nov;71(11):1063-5
16320210
Basic Clin Pharmacol Toxicol. 2009 Jun;104(6):463-9
19389047
J Cell Biochem. 2012 Nov;113(11):3476-85
22689083
PLoS One. 2012;7(5):e37205
22606351
Mol Cell Biol. 2014 Jul;34(13):2382-95
24732804
PLoS One. 2012;7(4):e34303
22558087
Cancer Res. 2007 Mar 15;67(6):2816-23
17363604
Cell. 2009 Jan 23;136(2):215-33
19167326
Clin Cancer Res. 2003 Dec 15;9(17):6371-80
14695137
J Natl Cancer Inst. 2012 Sep 19;104(18):1396-410
22972969
J Ethnopharmacol. 2000 Sep;72(1-2):43-6
10967452
Nature. 2001 Jul 5;412(6842):95-9
11452314
Int J Mol Sci. 2015 Jan 23;16(2):2530-58
25625511
Prev Med. 1981 Sep;10(5):614-22
6272265
BMC Cancer. 2015 Dec 16;15:974
26673922
Trends Cell Biol. 2004 Feb;14(2):70-7
15102438
Cancer Epidemiol Biomarkers Prev. 2008 Jul;17(7):1648-52
18628415
J Exp Clin Cancer Res. 2016 Jan 22;35:17
26795575
J Biol Chem. 2009 Aug 7;284(32):21412-24
19520853