NLK functions to maintain proliferation and stemness of NSCLC and is a target of metformin

被引:39
作者
Dong Suwei [1 ,2 ]
Zeng Liang [3 ]
Liu Zhimin [2 ]
Li Ruilei [2 ]
Zou Yingying [4 ]
Li Zhen [1 ,2 ]
Ge Chunlei [2 ]
Lai Zhangchao [2 ]
Xue Yuanbo [2 ]
Yang Jinyan [2 ]
Li Gaofeng [5 ]
Song Xin [1 ,2 ]
机构
[1] Southern Med Univ, Canc Res Inst, Guangzhou, Guangdong, Peoples R China
[2] Kunming Med Univ, Tumor Hosp Yunnan Prov, Affiliated Hosp 3, Dept Canc Biotherapy Ctr, Kunming, Yunnan, Peoples R China
[3] Hunan Tumor Hosp, Dept Pathol, Changsha, Hunan, Peoples R China
[4] Kunming Med Univ, Dept Pathol & Pathophysiol, Kunming, Yunnan, Peoples R China
[5] Kunming Med Univ, Tumor Hosp Yunnan Prov, Affiliated Hosp 3, Dept Thorac Surg, Kunming, Yunnan, Peoples R China
基金
中国国家自然科学基金; 国家高技术研究发展计划(863计划);
关键词
NLK; NSCLC; Proliferation; Metformin; Stemness; NEMO-LIKE KINASE; CYCLIN D1; IN-VITRO; CANCER; PROTEIN; PATHWAY; EXPRESSION; GROWTH; CELLS; DEGRADATION;
D O I
10.1186/s13045-015-0203-8
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Objective: Nemo-like kinase (NLK) is an evolutionarily conserved serine/threonine kinase that regulates the activity of a wide range of signal transduction pathways. Metformin, an oral antidiabetic drug, is used for cancer prevention. However, the significance and underlying mechanism of NLK and metformin in oncogenesis has not been fully elucidated. Here, we investigate a novel role of NLK and metformin in human non-small cell lung cancer (NSCLC). Materials and methods: NLK expression was analyzed in 121 NSCLCs and 92 normal lung tissue samples from benign pulmonary disease. Lentivirus vectors with NLK-shRNA were used to examine the effect of NLK on cell proliferation and tumorigenesis in vitro. Then, tumor xenograft mouse models revealed that NLK knockdown cells had a reduced ability for tumor formation compared with the control group in vivo. Multiple cell cycle regulator expression patterns induced by NLK silencing were examined by western blots in A549 cells. We also employed metformin to study its anti-cancer effects and mechanisms. Cancer stem cell property was checked by tumor sphere formation and markers including CD133, Nanog, c-Myc, and TLF4. Results: Immunohistochemical (IHC) analysis revealed that NLK expression was up-regulated in NSCLC cases (p < 0.001) and correlated with tumor T stage (p < 0.05). Silencing of NLK suppressed cell proliferation and tumorigenicity significantly in vitro and in vivo, which might be modulated by JUN family proteins. Furthermore, metformin selectively inhibits NLK expression and proliferation in NSCLC cells, but not immortalized noncancerous lung bronchial epithelial cells. In addition, both NLK knockdown and metformin treatment reduced the tumor sphere formation capacity and percentage of CD133+ cells. Accordingly, the expression level of stem cell markers (Nanog, c-Myc, and TLF4) were decreased significantly. Conclusion: NLK is critical for cancer cell cycle progression, and tumorigenesis in NSCLC, NLK knockdown, and metformin treatment inhibit cancer cell proliferation and stemness. Metformin inhibits NLK expression and might be a potential treatment strategy for NSCLC.
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页数:14
相关论文
共 47 条
[1]  
Anisimov VN, 2015, CURR DRUG TARGETS, V3, P3
[2]   Metformin Inhibits Cell Proliferation, Migration and Invasion by Attenuating CSC Function Mediated by Deregulating miRNAs in Pancreatic Cancer Cells [J].
Bao, Bin ;
Wang, Zhiwei ;
Ali, Shadan ;
Ahmad, Aamir ;
Azmi, Asfar S. ;
Sarkar, Sanila H. ;
Banerjee, Sanjeev ;
Kong, Dejuan ;
Li, Yiwei ;
Thakur, Shivam ;
Sarkar, Fazlul H. .
CANCER PREVENTION RESEARCH, 2012, 5 (03) :355-364
[3]   Nlk is a murine protein kinase related to Erk/MAP kinases and localized in the nucleus [J].
Brott, BK ;
Pinsky, BA ;
Erikson, RL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (03) :963-968
[4]   ROTATION OF PHOTORECEPTOR CLUSTERS IN THE DEVELOPING DROSOPHILA EYE REQUIRES THE NEMO GENE [J].
CHOI, KW ;
BENZER, S .
CELL, 1994, 78 (01) :125-136
[5]  
Doerfler W, 2001, ANN NY ACAD SCI, V945, P276
[6]   NLK functions to maintain proliferation and stemness of NSCLC and is a target of metformin [J].
Dong Suwei ;
Zeng Liang ;
Liu Zhimin ;
Li Ruilei ;
Zou Yingying ;
Li Zhen ;
Ge Chunlei ;
Lai Zhangchao ;
Xue Yuanbo ;
Yang Jinyan ;
Li Gaofeng ;
Song Xin .
JOURNAL OF HEMATOLOGY & ONCOLOGY, 2015, 8
[7]  
EDELMAN AM, 1987, ANNU REV BIOCHEM, V56, P567, DOI 10.1146/annurev.biochem.56.1.567
[8]   The American Joint Committee on Cancer: the 7th Edition of the AJCC Cancer Staging Manual and the Future of TNM [J].
Edge, Stephen B. ;
Compton, Carolyn C. .
ANNALS OF SURGICAL ONCOLOGY, 2010, 17 (06) :1471-1474
[9]   Nemo-Like Kinase Induces Apoptosis and Inhibits Androgen Receptor Signaling in Prostate Cancer Cells [J].
Emami, Katayoon H. ;
Brown, Lisha G. ;
Pitts, Tiffany E. M. ;
Sun, Xizhang ;
Vessella, Robert L. ;
Corey, Eva .
PROSTATE, 2009, 69 (14) :1481-1492
[10]   Metformin and reduced risk of cancer in diabetic patients [J].
Evans, JMM ;
Donnelly, LA ;
Emslie-Smith, AM ;
Alessi, DR ;
Morris, AD .
BMJ-BRITISH MEDICAL JOURNAL, 2005, 330 (7503) :1304-1305