Netrin‑1 interference potentiates epithelial‑to‑mesenchymal transition through the PI3K/AKT pathway under the hypoxic microenvironment conditions of non‑small cell lung cancer

  • Authors:
    • Xiayun Jin
    • Heqi Luan
    • Hua Chai
    • Lina Yan
    • Jing Zhang
    • Qi Wang
    • Lihua Cao
  • View Affiliations

  • Published online on: February 15, 2019     https://doi.org/10.3892/ijo.2019.4716
  • Pages: 1457-1465
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Abstract

Netrin‑1 is overexpressed in several types of cancer. However, whether netrin‑1 can potentiate hypoxia‑induced tumor progression in lung cancer has not been reported to date. Thus, the objective of the present study was to investigate whether netrin‑1 regulates cancer cell migration and invasion under hypoxic conditions in lung cancer and explore the underlying mechanism. A three‑dimensional microfluidic chip was used to observe real‑time changes in cancer cells, and cobalt chloride (CoCl2) was used to simulate a hypoxic microenvironment. Netrin‑1 siRNA was employed in the A549 and PC9 cell lines to downregulate the expression of netrin‑1. Microfluidic chip, wound healing and Transwell assays were used to examine cell migration and invasion. The expression levels of E‑cadherin and vimentin were detected by western blotting. The data demonstrated that netrin‑1 mediated epithelial‑to‑mesenchymal transition (EMT) of A549 and PC9 cells in vitro, which may be associated with the phosphoinositide 3 kinase/AKT pathway. This effect of netrin‑1 on the EMT was not observed in the normoxic microenvironment. In this retrospective study, netrin‑1 concentrations were evaluated in serum obtained from patients with non‑small cell lung cancer (NSCLC) and compared with healthy control samples by quantitative enzyme‑linked immunosorbent analysis. The serum concentration of netrin‑1 was found to be significantly higher in NSCLC patients compared with that in healthy donors. Taken together, the findings of the present study highlight a novel role for netrin‑1 in tumor development under hypoxia in NSCLC and provide further evidence for the use of netrin‑1 as a therapeutic target.

References

1 

Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI

2 

Shaikh D, Zhou Q, Chen T, Ibe JC, Raj JU and Zhou G: cAMP-dependent protein kinase is essential for hypoxia-mediated epithelial-mesenchymal transition, migration, and invasion in lung cancer cells. Cell Signal. 24:2396–2406. 2012. View Article : Google Scholar : PubMed/NCBI

3 

Gunther S, Ostheimer C, Stangl S, Specht HM, Mozes P, Jesinghaus M, Vordermark D, Combs SE, Peltz F, Jung MP, et al: Correlation of Hsp70 Serum Levels with Gross Tumor Volume and Composition of Lymphocyte Subpopulations in Patients with Squamous Cell and Adeno Non-Small Cell Lung Cancer. Front Immunol. 6:5562015. View Article : Google Scholar : PubMed/NCBI

4 

Foss KM, Sima C, Ugolini D, Neri M, Allen KE and Weiss GJ: miR-1254 and miR-574-5p: serum-based microRNA biomarkers for early-stage non-small cell lung cancer. J Thorac Oncol. 6:482–488. 2011. View Article : Google Scholar : PubMed/NCBI

5 

Wang S, Li E, Gao Y, Wang Y, Guo Z, He J, Zhang J, Gao Z and Wang Q: Study on invadopodia formation for lung carcinoma invasion with a microfluidic 3D culture device. PLoS One. 8:e564482013. View Article : Google Scholar : PubMed/NCBI

6 

Zhao M, Zhang Y, Zhang H, Wang S, Zhang M, Chen X, Wang H, Zeng G, Chen X, Liu G, et al: Hypoxia-induced cell stemness leads to drug resistance and poor prognosis in lung adenocar-cinoma. Lung Cancer. 87:98–106. 2015. View Article : Google Scholar

7 

Ye LY, Chen W, Bai XL, Xu XY, Zhang Q, Xia XF, Sun X, Li GG, Hu QD, Fu QH, et al: Hypoxia-Induced Epithelial-to-Mesenchymal Transition in Hepatocellular Carcinoma Induces an Immunosuppressive Tumor Microenvironment to Promote Metastasis. Cancer Res. 76:818–830. 2016. View Article : Google Scholar : PubMed/NCBI

8 

Zhang L, Huang G, Li X, Zhang Y, Jiang Y, Shen J, Liu J, Wang Q, Zhu J, Feng X, et al: Hypoxia induces epithelial-mesenchymal transition via activation of SNAI1 by hypoxia-inducible factor-1α in hepatocellular carcinoma. BMC Cancer. 13:1082013. View Article : Google Scholar

9 

Chen S, Chen JZ, Zhang JQ, Chen HX, Yan ML, Huang L, Tian YF, Chen YL and Wang YD: Hypoxia induces TWIST-activated epithelial-mesenchymal transition and proliferation of pancreatic cancer cells in vitro and in nude mice. Cancer Lett. 383:73–84. 2016. View Article : Google Scholar : PubMed/NCBI

10 

Masoud GN and Li W: HIF-1α pathway: Role, regulation and intervention for cancer therapy. Acta Pharm Sin B. 5:378–389. 2015. View Article : Google Scholar : PubMed/NCBI

11 

Vaupel P and Multhoff G: Accomplices of the Hypoxic Tumor Microenvironment Compromising Antitumor Immunity: Adenosine, Lactate, Acidosis, Vascular Endothelial Growth Factor, Potassium Ions, and Phosphatidylserine. Front Immunol. 8:18872017. View Article : Google Scholar

12 

Nakamura H, Ichikawa T, Nakasone S, Miyoshi T, Sugano M, Kojima M, Fujii S, Ochiai A, Kuwata T, Aokage K, et al: Abundant tumor promoting stromal cells in lung adenocarcinoma with hypoxic regions. Lung Cancer. 115:56–63. 2018. View Article : Google Scholar : PubMed/NCBI

13 

Dominici C, Moreno-Bravo JA, Puiggros SR, Rappeneau Q, Rama N, Vieugue P, Bernet A, Mehlen P and Chédotal A: Floor-plate-derived netrin-1 is dispensable for commissural axon guidance. Nature. 545:350–354. 2017. View Article : Google Scholar : PubMed/NCBI

14 

Xu K, Wu Z, Renier N, Antipenko A, Tzvetkova-Robev D, Xu Y, Minchenko M, Nardi-Dei V, Rajashankar KR, Himanen J, et al: Neural migration. Structures of netrin-1 bound to two receptors provide insight into its axon guidance mechanism. Science. 344:1275–1279. 2014. View Article : Google Scholar : PubMed/NCBI

15 

Dun XP and Parkinson DB: Role of Netrin-1 Signaling in Nerve Regeneration. Int J Mol Sci. 18:182017. View Article : Google Scholar

16 

Chen J, Du H, Zhang Y, Chen H, Zheng M, Lin P, Lan Q, Yuan Q, Lai Y, Pan X, et al: Netrin-1 Prevents Rat Primary Cortical Neurons from Apoptosis via the DCC/ERK Pathway. Front Cell Neurosci. 11:3872017. View Article : Google Scholar

17 

Bai L, Mei X, Wang Y, Yuan Y, Bi Y, Li G, Wang H, Yan P and Lv G: The Role of Netrin-1 in Improving Functional Recovery through Autophagy Stimulation Following Spinal Cord Injury in Rats. Front Cell Neurosci. 11:3502017. View Article : Google Scholar : PubMed/NCBI

18 

Ko SY, Blatch GL and Dass CR: Netrin-1 as a potential target for metastatic cancer: Focus on colorectal cancer. Cancer Metastasis Rev. 33:101–113. 2014. View Article : Google Scholar

19 

Lv J, Sun X, Ma J, Ma X, Zhang Y, Li F, Li Y and Zhao Z: Netrin-1 induces the migration of Schwann cells via p38 MAPK and PI3K-Akt signaling pathway mediated by the UNC5B receptor. Biochem Biophys Res Commun. 464:263–268. 2015. View Article : Google Scholar : PubMed/NCBI

20 

Akino T, Han X, Nakayama H, McNeish B, Zurakowski D, Mammoto A, Klagsbrun M and Smith E: Netrin-1 promotes medulloblastoma cell invasiveness and angiogenesis, and demonstrates elevated expression in tumor tissue and urine of patients with pediatric medulloblastoma. Cancer Res. 74:3716–3726. 2014. View Article : Google Scholar : PubMed/NCBI

21 

Grandin M, Mathot P, Devailly G, Bidet Y, Ghantous A, Favrot C, Gibert B, Gadot N, Puisieux I, Herceg Z, Delcros JG, Bernet A and Mehlen P: Inhibition of DNA methylation promotes breast tumor sensitivity to netrin-1 interference. 8:863–877. 2016.

22 

Wang H, Zhang B, Gu M, Li S, Chi Z and Hao L: Overexpression of the dependence receptor UNC5H4 inhibits cell migration and invasion, and triggers apoptosis in neuroblastoma cell. Tumour Biol. 35:5417–5425. 2014. View Article : Google Scholar : PubMed/NCBI

23 

Gibert B and Mehlen P: Dependence Receptors and Cancer: Addiction to Trophic Ligands. Cancer Res. 75:5171–5175. 2015. View Article : Google Scholar : PubMed/NCBI

24 

Zhang Y, Wang B, Chen X, Li W and Dong P: AGO2 involves the malignant phenotypes and FAK/PI3K/AKT signaling pathway in hypopharyngeal-derived FaDu cells. Oncotarget. 8:54735–54746. 2017.PubMed/NCBI

25 

Zhang PF, Li KS, Shen YH, Gao PT, Dong ZR, Cai JB, Zhang C, Huang XY, Tian MX, Hu ZQ, et al: Galectin-1 induces hepato-cellular carcinoma EMT and sorafenib resistance by activating FAK/PI3K/AKT signaling. Cell Death Dis. 7:e22012016. View Article : Google Scholar

26 

Yang X, Li S, Zhong J, Zhang W, Hua X, Li B and Sun H: CD151 mediates netrin-1-induced angiogenesis through the Src-FAK-Paxillin pathway. J Cell Mol Med. 21:72–80. 2017. View Article : Google Scholar

27 

Song J, Zhang Y, Zhang C, Du X, Guo Z, Kuang Y, Wang Y, Wu P, Zou K, Zou L, et al: A microfluidic device for studying chemotaxis mechanism of bacterial cancer targeting. Sci Rep. 8:63942018. View Article : Google Scholar : PubMed/NCBI

28 

Torre LA, Siegel RL and Jemal A: Lung Cancer Statistics. Adv Exp Med Biol. 893:1–19. 2016. View Article : Google Scholar

29 

Quéré G, Descourt R, Robinet G, Autret S, Raguenes O, Fercot B, Alemany P, Uguen A, Férec C, Quintin-Roué I, et al: Mutational status of synchronous and metachronous tumor samples in patients with metastatic non-small-cell lung cancer. BMC Cancer. 16:2102016. View Article : Google Scholar : PubMed/NCBI

30 

Chu CY, Jin YT, Zhang W, Yu J, Yang HP, Wang HY, Zhang ZJ, Liu XP and Zou Q: CA IX is upregulated in CoCl2-induced hypoxia and associated with cell invasive potential and a poor prognosis of breast cancer. Int J Oncol. 48:271–280. 2016. View Article : Google Scholar

31 

Papanastasiou AD, Pampalakis G, Katsaros D and Sotiropoulou G: Netrin-1 overexpression is predictive of ovarian malignancies. Oncotarget. 2:363–367. 2011. View Article : Google Scholar : PubMed/NCBI

32 

Harter PN, Zinke J, Scholz A, Tichy J, Zachskorn C, Kvasnicka HM, Goeppert B, Delloye-Bourgeois C, Hattingen E, Senft C, et al: Netrin-1 expression is an independent prognostic factor for poor patient survival in brain metastases. PLoS One. 9:e923112014. View Article : Google Scholar : PubMed/NCBI

33 

An XZ, Zhao ZG, Luo YX, Zhang R, Tang XQ, Hao D, Zhao X, Lv X and Liu D: Netrin-1 suppresses the MEK/ERK pathway and ITGB4 in pancreatic cancer. Oncotarget. 7:24719–24733. 2016. View Article : Google Scholar : PubMed/NCBI

34 

Han P, Fu Y, Liu J, Wang Y, He J, Gong J, Li M, Tan Q, Li D, Luo Y, et al: Netrin-1 promotes cell migration and invasion by down-regulation of BVES expression in human hepatocellular carcinoma. Am J Cancer Res. 5:1396–1409. 2015.PubMed/NCBI

35 

Ducarouge B, Delcros JG, Abès R, Goldschneider D, Gibert B, Blachier J, Neves D, Mehlen P, Bernet A and Depil S: Abstract 2921: Preclinical characteristics of NP137, a first-in-class monoclonal antibody directed against netrin-1 and inducing dependence receptors-mediated cell death. Cancer Res. 75(Suppl): 2921. 2015. View Article : Google Scholar

36 

Fujiki K, Inamura H, Miyayama T and Matsuoka M: Involvement of Notch1 signaling in malignant progression of A549 cells subjected to prolonged cadmium exposure. J Biol Chem. 292:7942–7953. 2017. View Article : Google Scholar : PubMed/NCBI

37 

Hong CF, Chen WY and Wu CW: Upregulation of Wnt signaling under hypoxia promotes lung cancer progression. Oncol Rep. 38:1706–1714. 2017. View Article : Google Scholar : PubMed/NCBI

38 

Zhou Z, Wang S, Song C and Hu Z: Paeoniflorin prevents hypoxia-induced epithelial-mesenchymal transition in human breast cancer cells. OncoTargets Ther. 9:2511–2518. 2016. View Article : Google Scholar

39 

Zhang X, Cui P, Ding B, Guo Y, Han K, Li J, Chen H and Zhang W: Netrin-1 elicits metastatic potential of non-small cell lung carcinoma cell by enhancing cell invasion, migration and vasculogenic mimicry via EMT induction. Cancer Gene Ther. 25:18–26. 2018. View Article : Google Scholar

40 

Wang H, Zhang C, Xu L, Zang K, Ning Z, Jiang F, Chi H, Zhu X and Meng Z: Bufalin suppresses hepatocellular carcinoma invasion and metastasis by targeting HIF-1α via the PI3K/AKT/mTOR pathway. Oncotarget. 7:20193–20208. 2016.PubMed/NCBI

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Copy and paste a formatted citation
APA
Jin, X., Luan, H., Chai, H., Yan, L., Zhang, J., Wang, Q., & Cao, L. (2019). Netrin‑1 interference potentiates epithelial‑to‑mesenchymal transition through the PI3K/AKT pathway under the hypoxic microenvironment conditions of non‑small cell lung cancer. International Journal of Oncology, 54, 1457-1465. https://doi.org/10.3892/ijo.2019.4716
MLA
Jin, X., Luan, H., Chai, H., Yan, L., Zhang, J., Wang, Q., Cao, L."Netrin‑1 interference potentiates epithelial‑to‑mesenchymal transition through the PI3K/AKT pathway under the hypoxic microenvironment conditions of non‑small cell lung cancer". International Journal of Oncology 54.4 (2019): 1457-1465.
Chicago
Jin, X., Luan, H., Chai, H., Yan, L., Zhang, J., Wang, Q., Cao, L."Netrin‑1 interference potentiates epithelial‑to‑mesenchymal transition through the PI3K/AKT pathway under the hypoxic microenvironment conditions of non‑small cell lung cancer". International Journal of Oncology 54, no. 4 (2019): 1457-1465. https://doi.org/10.3892/ijo.2019.4716