Silencing NACK by siRNA inhibits tumorigenesis in non-small cell lung cancer via targeting Notch1 signaling pathway

Kong,Ranran; Feng,Jie; Ma,Yuefeng; Zhou,Bin; Li,Shaomin; Zhang,Wei; Jiang,Jiantao; Zhang,Jin; Qiao,Zhe; Zhang,Ting; Zang,Quanjin; He,Xijing

doi:10.3892/or.2016.4552

Silencing NACK by siRNA inhibits tumorigenesis in non-small cell lung cancer via targeting Notch1 signaling pathway

Authors:
- Ranran Kong
- Jie Feng
- Yuefeng Ma
- Bin Zhou
- Shaomin Li
- Wei Zhang
- Jiantao Jiang
- Jin Zhang
- Zhe Qiao
- Ting Zhang
- Quanjin Zang
- Xijing He
View Affiliations

Affiliations: Department of Thoracic Surgery, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China, Department of Nephrology, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Second Department of Orthopedics, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China
Published online on: January 13, 2016 https://doi.org/10.3892/or.2016.4552
Pages: 2306-2314

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Non-small cell lung cancer (NSCLC) is the most common type of lung tumor with poor prognosis, in which the Notch signaling pathway plays an important role. Notch activation complex kinase (NACK) has been reported both as a co-activator and a target gene of the Notch pathway. However, the molecular mechanism of NACK in NSCLC still remains unknown. In this study, the expression of NACK was analyzed in 35 paired NSCLC tumor samples and 2 NSCLC cell lines. MTT assay, cell migration assay, cell invasion assay, flow cytometry assay, and xenograft model were employed to detect the effect of NACK knockdown on the cell proliferation, metastasis, invasion and apoptosis of NSCLC. The relationship between NACK and Notch1 signaling pathway in NSCLC cells was assessed by western blot and luciferase reporter assay. We found that the expression of NACK in the NSCLC tissues and lung cancer cells were significantly increased. High level of NACK expression is remarkable associated with tumor differentiation, lymphatic metastasis, clinical stage and poor survival prognosis. The interference of NACK significantly inhibited cell proliferation, invasion and metastasis through inducing apoptosis in NSCLC cells. The transcriptional activity of related Notch1 target genes were significantly suppressed resulting from NACK knockdown. This study demonstrates that interference of NACK inhibits NSCLC progression through Notch1 signaling pathway and targeting NACK may be an effective approach for NSCLC therapy.

View Figures

View References

1	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar : PubMed/NCBI
2	Nguyen KS, Neal JW and Wakelee H: Review of the current targeted therapies for non-small-cell lung cancer. World J Clin Oncol. 5:576–587. 2014. View Article : Google Scholar : PubMed/NCBI
3	Boolell V, Alamgeer M, Watkins DN and Ganju V: The evolution of therapies in non-small cell lung cancer. Cancers (Basel). 7:1815–1846. 2015. View Article : Google Scholar
4	Rovigatti U: Cancer modelling in the NGS era - Part I: Emerging technology and initial modelling. Crit Rev Oncol Hematol. 96:274–307. 2015. View Article : Google Scholar : PubMed/NCBI
5	Gottfried M, Keizman D, Mishaeli M and Rabinovich NM: The current approach to advanced lung cancer. Harefuah. 154:521–540. 2015.In Hebrew.
6	Wang Z, Li Y, Banerjee S and Sarkar FH: Emerging role of Notch in stem cells and cancer. Cancer Lett. 279:8–12. 2009. View Article : Google Scholar :
7	Kristoffersen K, Villingshøj M, Poulsen HS and Stockhausen MT: Level of Notch activation determines the effect on growth and stem cell-like features in glioblastoma multiforme neurosphere cultures. Cancer Biol Ther. 14:625–637. 2013. View Article : Google Scholar : PubMed/NCBI
8	Jang MS, Miao H, Carlesso N, Shelly L, Zlobin A, Darack N, Qin JZ, Nickoloff BJ and Miele L: Notch-1 regulates cell death independently of differentiation in murine erythroleukemia cells through multiple apoptosis and cell cycle pathways. J Cell Physiol. 199:418–433. 2004. View Article : Google Scholar : PubMed/NCBI
9	Li L, Zhao F, Lu J, Li T, Yang H, Wu C and Liu Y: Notch-1 signaling promotes the malignant features of human breast cancer through NF-κB activation. PLoS One. 9:e959122014. View Article : Google Scholar
10	Leong KG and Karsan A: Recent insights into the role of Notch signaling in tumorigenesis. Blood. 107:2223–2233. 2006. View Article : Google Scholar
11	Leong KG and Gao WQ: The Notch pathway in prostate development and cancer. Differentiation. 76:699–716. 2008. View Article : Google Scholar : PubMed/NCBI
12	Miele L and Osborne B: Arbiter of differentiation and death: Notch signaling meets apoptosis. J Cell Physiol. 181:393–409. 1999. View Article : Google Scholar : PubMed/NCBI
13	Wang Z, Li Y, Banerjee S and Sarkar FH: Exploitation of the Notch signaling pathway as a novel target for cancer therapy. Anticancer Res. 28:3621–3630. 2008.
14	Yin L, Velazquez OC and Liu ZJ: Notch signaling: Emerging molecular targets for cancer therapy. Biochem Pharmacol. 80:690–701. 2010. View Article : Google Scholar : PubMed/NCBI
15	Weaver KL, Alves-Guerra MC, Jin K, Wang Z, Han X, Ranganathan P, Zhu X, DaSilva T, Liu W, Ratti F, et al: NACK is an integral component of the Notch transcriptional activation complex and is critical for development and tumorigenesis. Cancer Res. 74:4741–4751. 2014. View Article : Google Scholar : PubMed/NCBI
16	Androutsellis-Theotokis A, Leker RR, Soldner F, Hoeppner DJ, Ravin R, Poser SW, Rueger MA, Bae SK, Kittappa R and McKay RD: Notch signalling regulates stem cell numbers in vitro and in vivo. Nature. 442:823–826. 2006. View Article : Google Scholar : PubMed/NCBI
17	Bi P and Kuang S: Notch signaling as a novel regulator of metabolism. Trends Endocrinol Metab. 26:248–255. 2015. View Article : Google Scholar : PubMed/NCBI
18	Guruharsha KG, Kankel MW and Artavanis-Tsakonas S: The Notch signalling system: Recent insights into the complexity of a conserved pathway. Nat Rev Genet. 13:654–666. 2012. View Article : Google Scholar : PubMed/NCBI
19	Galluzzo P and Bocchetta M: Notch signaling in lung cancer. Expert Rev Anticancer Ther. 11:533–540. 2011. View Article : Google Scholar : PubMed/NCBI
20	Nickoloff BJ, Osborne BA and Miele L: Notch signaling as a therapeutic target in cancer: A new approach to the development of cell fate modifying agents. Oncogene. 22:6598–6608. 2003. View Article : Google Scholar : PubMed/NCBI
21	Ranganathan P, Weaver KL and Capobianco AJ: Notch signalling in solid tumours: A little bit of everything but not all the time. Nat Rev Cancer. 11:338–351. 2011. View Article : Google Scholar : PubMed/NCBI
22	Takebe N, Harris PJ, Warren RQ and Ivy SP: Targeting cancer stem cells by inhibiting Wnt, Notch, and Hedgehog pathways. Nat Rev Clin Oncol. 8:97–106. 2011. View Article : Google Scholar
23	Wael H, Yoshida R, Kudoh S, Hasegawa K, Niimori-Kita K and Ito T: Notch1 signaling controls cell proliferation, apoptosis and differentiation in lung carcinoma. Lung Cancer. 85:131–140. 2014. View Article : Google Scholar : PubMed/NCBI
24	Jiang X, Zhou JH, Deng ZH, Qu XH, Jiang HY and Liu Y: Expression and significance of Notch1, Jagged1 and VEGF in human non-small cell lung cancer. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 32:1031–1036. 2007.In Chinese.
25	Chen Y, De Marco MA, Graziani I, Gazdar AF, Strack PR, Miele L and Bocchetta M: Oxygen concentration determines the biological effects of NOTCH-1 signaling in adenocarcinoma of the lung. Cancer Res. 67:7954–7959. 2007. View Article : Google Scholar : PubMed/NCBI
26	Baumgart A, Seidl S, Vlachou P, Michel L, Mitova N, Schatz N, Specht K, Koch I, Schuster T, Grundler R, et al: ADAM17 regulates epidermal growth factor receptor expression through the activation of Notch1 in non-small cell lung cancer. Cancer Res. 70:5368–5378. 2010. View Article : Google Scholar : PubMed/NCBI
27	Ji X, Wang Z, Geamanu A, Sarkar FH and Gupta SV: Inhibition of cell growth and induction of apoptosis in non-small cell lung cancer cells by delta-tocotrienol is associated with notch-1 down-regulation. J Cell Biochem. 112:2773–2783. 2011. View Article : Google Scholar : PubMed/NCBI
28	Eliasz S, Liang S, Chen Y, De Marco MA, Machek O, Skucha S, Miele L and Bocchetta M: Notch-1 stimulates survival of lung adenocarcinoma cells during hypoxia by activating the IGF-1R pathway. Oncogene. 29:2488–2498. 2010. View Article : Google Scholar : PubMed/NCBI
29	Chen Y, Li D, Liu H, Xu H, Zheng H, Qian F, Li W, Zhao C, Wang Z and Wang X: Notch-1 signaling facilitates survivin expression in human non-small cell lung cancer cells. Cancer Biol Ther. 11:14–21. 2011. View Article : Google Scholar
30	Manning G, Whyte DB, Martinez R, Hunter T and Sudarsanam S: The protein kinase complement of the human genome. Science. 298:1912–1934. 2002. View Article : Google Scholar : PubMed/NCBI
31	Tanaka H, Katoh H and Negishi M: Pragmin, a novel effector of Rnd2 GTPase, stimulates RhoA activity. J Biol Chem. 281:10355–10364. 2006. View Article : Google Scholar : PubMed/NCBI
32	Nguyen TA and Fruehauf JH: Transkingdom RNA interference (tkRNAi): A novel method to induce therapeutic gene silencing. Methods Mol Biol. 514:27–34. 2009. View Article : Google Scholar
33	Geng J, Li X, Zhou Z, Wu CL, Dai M and Bai X: EZH2 promotes tumor progression via regulating VEGF-A/AKT signaling in non-small cell lung cancer. Cancer Lett. 359:275–287. 2015. View Article : Google Scholar : PubMed/NCBI
34	Koff JL, Ramachandiran S and Bernal-Mizrachi L: A time to kill: Targeting apoptosis in cancer. Int J Mol Sci. 16:2942–2955. 2015. View Article : Google Scholar : PubMed/NCBI
35	Drosopoulos K and Pintzas A: Multifaceted targeting in cancer: The recent cell death players meet the usual oncogene suspects. Expert Opin Ther Targets. 11:641–659. 2007. View Article : Google Scholar : PubMed/NCBI
36	Taylor K, Micha D, Ranson M and Dive C: Recent advances in targeting regulators of apoptosis in cancer cells for therapeutic gain. Expert Opin Investig Drugs. 15:669–690. 2006. View Article : Google Scholar : PubMed/NCBI
37	Fischer A and Gessler M: Delta-Notch - and then? Protein interactions and proposed modes of repression by Hes and Hey bHLH factors. Nucleic Acids Res. 35:4583–4596. 2007. View Article : Google Scholar :
38	Nakagawa O, McFadden DG, Nakagawa M, Yanagisawa H, Hu T, Srivastava D and Olson EN: Members of the HRT family of basic helix-loop-helix proteins act as transcriptional repressors downstream of Notch signaling. Proc Natl Acad Sci USA. 97:13655–13660. 2000. View Article : Google Scholar : PubMed/NCBI
39	Iso T, Kedes L and Hamamori Y: HES and HERP families: Multiple effectors of the Notch signaling pathway. J Cell Physiol. 194:237–255. 2003. View Article : Google Scholar : PubMed/NCBI
40	Al-Hussaini H, Subramanyam D, Reedijk M and Sridhar SS: Notch signaling pathway as a therapeutic target in breast cancer. Mol Cancer Ther. 10:9–15. 2011. View Article : Google Scholar
41	Candy PA, Phillips MR, Redfern AD, Colley SM, Davidson JA, Stuart LM, Wood BA, Zeps N and Leedman PJ: Notch-induced transcription factors are predictive of survival and 5-fluorouracil response in colorectal cancer patients. Br J Cancer. 109:1023–1030. 2013. View Article : Google Scholar : PubMed/NCBI