SPINK6 Promotes Metastasis of Nasopharyngeal Carcinoma via Binding and Activation of Epithelial Growth Factor Receptor.
Nasopharyngeal carcinoma has the highest rate of metastasis among head and neck cancers, and distant metastasis is the major reason for treatment failure. The underlying molecular mechanisms of nasopharyngeal carcinoma metastasis are not fully understood. Here, we report the identification of serine protease inhibitor Kazal-type 6 (SPINK6) as a functional regulator of nasopharyngeal carcinoma metastasis via EGFR signaling. SPINK6 mRNA was upregulated in tumor and highly metastatic nasopharyngeal carcinoma cells. Immunohistochemical staining of 534 nasopharyngeal carcinomas revealed elevated SPINK6 expression as an independent unfavorable prognostic factor for overall, disease-free, and distant metastasis-free survival. Ectopic SPINK6 expression promoted in vitro migration and invasion as well as in vivo lymph node metastasis and liver metastasis of nasopharyngeal carcinoma cells, whereas silencing SPINK6 exhibited opposing effects. SPINK6 enhanced epithelial-mesenchymal transition by activating EGFR and the downstream AKT pathway. Inhibition of EGFR with a neutralizing antibody or erlotinib reversed SPINK6-induced nasopharyngeal carcinoma cell migration and invasion. Erlotinib also inhibited SPINK6-induced metastasis in vivo Notably, SPINK6 bound to the EGFR extracellular domain independent of serine protease-inhibitory activity. Overall, our results identified a novel EGFR-activating mechanism in which SPINK6 has a critical role in promoting nasopharyngeal carcinoma metastasis, with possible implications as a prognostic indicator in nasopharyngeal carcinoma patients. Cancer Res; 77(2); 579-89. ©2016 AACR.
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Li-Sheng
LS
Zheng
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Jun-Ping
JP
Yang
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Pathology, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Yun
Y
Cao
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Li-Xia
LX
Peng
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Rui
R
Sun
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Ping
P
Xie
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Radiotherapy Department, Affiliated Cancer Hospital of Guangzhou Medical University, Guangzhou, China.
Meng-Yao
MY
Wang
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Dong-Fang
DF
Meng
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Dong-Hua
DH
Luo
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Xiong
X
Zou
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Ming-Yuan
MY
Chen
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Hai-Qiang
HQ
Mai
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Ling
L
Guo
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Xiang
X
Guo
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.|Department of Molecular Diagnostics, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Jian-Yong
JY
Shao
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Bi-Jun
BJ
Huang
Department of Pathology, University of Texas MD Anderson Cancer Center, Houston, Texas.|Department of Cancer Biology, Comprehensive Cancer Center of Wake Forest Baptist Medical Center, Winston-Salem, North Carolina.
Wei
W
Zhang
State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China. qianchn@sysucc.org.cn.|Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Chao-Nan
CN
Qian
D002277:Carcinoma / Q000378:metabolism / Q000401:mortality / Q000473:pathology*; D002465:Cell Movement / Q000502:physiology; D018572:Disease-Free Survival; D058750:Epithelial-Mesenchymal Transition / Q000502:physiology; D066246:ErbB Receptors / Q000378:metabolism*; D005260:Female; D064593:Heterografts; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D053208:Kaplan-Meier Estimate; D008207:Lymphatic Metastasis / Q000473:pathology; D008297:Male; D000077274:Nasopharyngeal Carcinoma; D009303:Nasopharyngeal Neoplasms / Q000378:metabolism / Q000401:mortality / Q000473:pathology*; D009361:Neoplasm Invasiveness / Q000473:pathology; D011379:Prognosis; D016016:Proportional Hazards Models; D053491:Proteinase Inhibitory Proteins, Secretory / Q000378:metabolism*; D000074162:Serine Peptidase Inhibitors, Kazal Type; D046888:Tissue Array Analysis
D053491:Proteinase Inhibitory Proteins, Secretory; C553928:SPINK6 protein, human; D000074162:Serine Peptidase Inhibitors, Kazal Type; C512478:EGFR protein, human; D066246:ErbB Receptors
Prostate. 2014 May;74(7):689-701
24619958
Dis Markers. 2015;2015:762128
25999660
Exp Anim. 2011;60(5):433-44
22041280
J Biol Chem. 2009 Nov 6;284(45):30897-906
19717562
Biochem Biophys Res Commun. 2016 Feb 26;471(1):103-8
26828269
J Cancer Res Clin Oncol. 2007 Feb;133(2):71-81
16941191
BMC Cancer. 2010 Sep 17;10:498
20849596
Carcinogenesis. 2012 Jul;33(7):1302-9
22610073
Nature. 2001 Mar 1;410(6824):50-6
11242036
Biomark Med. 2015;9(10):997-1010
26441207
Cancer Res. 2006 Nov 1;66(21):10365-76
17062557
FEBS Lett. 2011 Feb 18;585(4):591-5
21255571
Head Neck. 2003 Oct;25(10):864-72
12966511
Head Neck. 2014 Oct;36(10):1508-16
23996630
J Invest Dermatol. 2013 May;133(5):1170-7
23303447
Oncotarget. 2015 Oct 6;6(30):29240-53
26335051
Int J Radiat Oncol Biol Phys. 2004 May 1;59(1):11-20
15093894
Apoptosis. 2008 Apr;13(4):483-94
18347987
Biol Chem. 2012 Apr;393(5):413-20
22505523
Oncogene. 2008 Jul 3;27(29):4024-33
18317448
FEBS Open Bio. 2014 Jan 13;4:128-40
24490137
Cell. 2002 Sep 20;110(6):763-73
12297049
Chin J Cancer. 2015 May 14;34(8):350-7
26058679
Sci Transl Med. 2011 Mar 2;3(72):72ra17
21368222
Future Oncol. 2010 Feb;6(2):269-85
20146586
Ai Zheng. 2002 Feb;21(2):158-62
12479066
Cancer Lett. 2010 Apr 1;290(1):87-95
19796867
Nat Genet. 2014 Aug;46(8):866-71
24952746
Eur J Cancer. 2010 Jul;46(11):1967-78
20451372
Carcinogenesis. 2007 Nov;28(11):2274-81
17602171
PLoS One. 2009 Aug 06;4(8):e6529
19657394
Expert Rev Mol Med. 2007 May 04;9(12):1-24
17477889
Mol Cell. 2003 Feb;11(2):507-17
12620237
Pharmacogenet Genomics. 2006 Jan;16(1):73-4
16344724
Cancer Res. 2011 Apr 15;71(8):3162-72
21289131
Oncotarget. 2015 Apr 30;6(12):10239-52
25823923
Jpn J Cancer Res. 1999 Dec;90(12):1285-92
10665644
Cancer Epidemiol Biomarkers Prev. 2006 Oct;15(10):1765-77
17035381
Ai Zheng. 2008 May;27(5):449-54
18479591
Cell. 2009 Nov 25;139(5):871-90
19945376
Oncotarget. 2014 May 15;5(9):2390-403
24158494
Oncotarget. 2016 Mar 29;7(13):15632-47
26701209
J Biol Chem. 2010 Oct 15;285(42):32174-81
20667819
Lancet. 2016 Mar 5;387(10022):1012-1024
26321262
J Invest Dermatol. 2011 Nov;131(11):2169-73
21997416
Hum Pathol. 2007 Jan;38(1):120-33
16996564
J Clin Oncol. 2015 Oct 10;33(29):3356-64
26351355
Biol Chem. 2010 Apr;391(4):299-310
20180639
Biol Chem. 2010 May;391(5):505-11
20302518
Peptides. 2011 Jun;32(6):1187-92
21439340