Downregulation of LRIG2 expression inhibits angiogenesis of glioma via EGFR/VEGF‑A pathway

Yang,Hong‑Kuan; Chen,Hao; Mao,Feng; Xiao,Qun‑Gen; Xie,Rui‑Fan; Lei,Ting

doi:10.3892/ol.2017.6671

Downregulation of LRIG2 expression inhibits angiogenesis of glioma via EGFR/VEGF‑A pathway

Authors:
- Hong‑Kuan Yang
- Hao Chen
- Feng Mao
- Qun‑Gen Xiao
- Rui‑Fan Xie
- Ting Lei
View Affiliations

Affiliations: Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China
Published online on: July 26, 2017 https://doi.org/10.3892/ol.2017.6671
Pages: 4021-4028
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Active angiogenesis is the basic pathological feature of glioma. Tumor angiogenesis is involved in vascular endothelial cell migration to the tumor tissue and in the formation of tube‑like structures. The present study aimed to investigate the role of leucine‑rich repeats and immunoglobulin‑like domains 2 (LRIG2) in glioma angiogenesis. Glioma (n=50) and normal brain (n=20) tissue samples were collected from patients to detect the expression of LRIG2, epidermal growth factor receptor (EGFR), vascular endothelial growth factor A (VEGF‑A), and cluster of differentiation 31 (CD31) using immunohistochemistry. In addition, the association between the expression of LRIG2 in glioma tissue and the microvessel density (MVD) was analyzed. In vitro, the expression of LRIG2 in human glioma U87 and U251 cell lines was knocked down. Subsequently, cell migration and tube formation assays of human umbilical vein endothelial cells (HUVECs) were performed using a coculture system. The protein expression levels of LRIG2, EGFR, phosphorylated‑EGFR and VEGF‑A were determined using western blotting. The results demonstrated that the expression levels of LRIG2, EGFR, VEGF‑A and CD31 were highly upregulated in glioma tissue samples. Furthermore, LRIG2 expression in glioma tissue samples was signiﬁcantly correlated with the MVD. In vitro, the downregulation of LRIG2 inhibited HUVEC migration and tube formation induced by coculture with glioma cells. The downregulation of LRIG2 resulted in decreased expression of EGFR and VEGF‑A. The effects of the LRIG2 knockdown were reversed following EGF treatment. These ﬁndings suggest that LRIG2 is a potential target for the inhibition of glioma angiogenesis, which is possibly mediated via the EGFR/VEGF‑A signaling pathway.

View Figures

View References

1	Wen PY and Reardon DA: Neuro-oncology in 2015: Progress in glioma diagnosis, classification and treatment. Nat Rev Neurol. 12:69–70. 2016. View Article : Google Scholar : PubMed/NCBI
2	Malla RR, Gopinath S, Gondi CS, Alapati K, Dinh DH, Gujrati M and Rao JS: Cathepsin B and uPAR knockdown inhibits tumor-induced angiogenesis by modulating VEGF expression in glioma. Cancer Gene Ther. 18:419–434. 2011. View Article : Google Scholar : PubMed/NCBI
3	Babykutty S SPPJNR, Kumar MA, Nair MS, Srinivas P and Gopala S: Nimbolide retards tumor cell migration, invasion, and angiogenesis by downregulating MMP-2/9 expression via inhibiting ERK1/2 and reducing DNA-binding activity of NF-κB in colon cancer cells. Mol Carcinog. 51:475–490. 2012. View Article : Google Scholar : PubMed/NCBI
4	Petrillo M, Borriello M, Fuoco G, Legge F, Iannone V and Ferrandina G: Novel VEGF-independent strategies targeting tumor vasculature: Clinical aspects. Curr Pharm Des. 18:2702–2712. 2012. View Article : Google Scholar : PubMed/NCBI
5	Reardon DA, Turner S, Peters KB, Desjardins A, Gururangan S, Sampson JH, McLendon RE, Herndon JE II, Jones LW, Kirkpatrick JP, et al: A review of VEGF/VEGFR-targeted therapeutics for recurrent glioblastoma. J Natl Compr Canc Netw. 9:414–427. 2011. View Article : Google Scholar : PubMed/NCBI
6	Cea V, Sala C and Verpelli C: Antiangiogenic therapy for glioma. J Signal Transduct. 2012:4830402012. View Article : Google Scholar : PubMed/NCBI
7	Casaletto JB and McClatchey AI: Spatial regulation of receptor tyrosine kinases in development and cancer. Nat Rev Cancer. 12:387–400. 2012. View Article : Google Scholar : PubMed/NCBI
8	Larsen AK, Ouaret D, El Ouadrani K and Petitprez A: Targeting EGFR and VEGF(R) pathway cross-talk in tumor survival and angiogenesis. Pharmacol Ther. 131:80–90. 2011. View Article : Google Scholar : PubMed/NCBI
9	Malik U and Javed A: LRIGs: A prognostically significant family with emerging therapeutic competence against cancers. Curr Cancer Drug Targets. 17:3–16. 2017. View Article : Google Scholar : PubMed/NCBI
10	van Erp S, van den Heuvel DM, Fujita Y, Robinson RA, Hellemons AJ, Adolfs Y, Van Battum EY, Blokhuis AM, Kuijpers M, Demmers JA, et al: Lrig2 negatively regulates ectodomain shedding of Axon guidance receptors by ADAM proteases. Dev Cell. 35:537–552. 2015. View Article : Google Scholar : PubMed/NCBI
11	Guo D, Nilsson J, Haapasalo H, Raheem O, Bergenheim T, Hedman H and Henriksson R: Perinuclear leucine-rich repeats and immunoglobulin-like domain proteins (LRIG1-3) as prognostic indicators in astrocytic tumors. Acta Neuropathol. 111:238–246. 2006. View Article : Google Scholar : PubMed/NCBI
12	Lindström AK, Asplund A and Hellberg D: Correlation between LRIG1 and LRIG2 expressions and expression of 11 tumor markers, with special reference to tumor suppressors, in CIN and normal cervical epithelium. Gynecol Oncol. 122:372–376. 2011. View Article : Google Scholar : PubMed/NCBI
13	Powell AE, Wang Y, Li Y, Poulin EJ, Means AL, Washington MK, Higginbotham JN, Juchheim A, Prasad N, Levy SE, et al: The pan-ErbB negative regulator Lrig1 is an intestinal stem cell marker that functions as a tumor suppressor. Cell. 149:146–158. 2012. View Article : Google Scholar : PubMed/NCBI
14	Wong VW, Stange DE, Page ME, Buczacki S, Wabik A, Itami S, van de Wetering M, Poulsom R, Wright NA, Trotter MW, et al: Lrig1 controls intestinal stem-cell homeostasis by negative regulation of ErbB signalling. Nat Cell Biol. 14:401–408. 2012. View Article : Google Scholar : PubMed/NCBI
15	Lu L, Teixeira VH, Yuan Z, Graham TA, Endesfelder D, Kolluri K, Al-Juffali N, Hamilton N, Nicholson AG, Falzon M, et al: LRIG1 regulates cadherin-dependent contact inhibition directing epithelial homeostasis and pre-invasive squamous cell carcinoma development. J Pathol. 229:608–620. 2013. View Article : Google Scholar : PubMed/NCBI
16	Bai L, McEachern D, Yang CY, Lu J, Sun H and Wang S: LRIG1 modulates cancer cell sensitivity to Smac mimetics by regulating TNFα expression and receptor tyrosine kinase signaling. Cancer Res. 72:1229–1238. 2012. View Article : Google Scholar : PubMed/NCBI
17	Hedman H, Lindström AK, Tot T, Stendahl U, Henriksson R and Hellberg D: LRIG2 in contrast to LRIG1 predicts poor survival in early-stage squamous cell carcinoma of the uterine cervix. Acta Oncol. 49:812–815. 2010. View Article : Google Scholar : PubMed/NCBI
18	Holmlund C, Haapasalo H, Yi W, Raheem O, Brännström T, Bragge H, Henriksson R and Hedman H: Cytoplasmic LRIG2 expression is associated with poor oligodendroglioma patient survival. Neuropathology. 29:242–247. 2009. View Article : Google Scholar : PubMed/NCBI
19	Wang B, Han L, Chen R, Cai M, Han F, Lei T and Guo D: Downregulation of LRIG2 expression by RNA interference inhibits glioblastoma cell (GL15) growth, causes cell cycle redistribution, increases cell apoptosis and enhances cell adhesion and invasion in vitro. Cancer Biol Ther. 8:1018–1023. 2009. View Article : Google Scholar : PubMed/NCBI
20	Remmele W and Stegner HE: Recommendation for uniform definition of an immunoreactive score (IRS) for immunohistochemical estrogen receptor detection (ER-ICA) in breast cancer tissue. Pathologe. 8:138–140. 1987.(In German). PubMed/NCBI
21	Tai YT, Acharya C, An G, Moschetta M, Zhong MY, Feng X, Cea M, Cagnetta A, Wen K, van Eenennaam H, et al: APRIL and BCMA promote human multiple myeloma growth and immunosuppression in the bone marrow microenvironment. Blood. 127:3225–3236. 2016. View Article : Google Scholar : PubMed/NCBI
22	Wang G, Wu J and Song H: LRIG2 expression and prognosis in non-small cell lung cancer. Oncol Lett. 8:667–672. 2014.PubMed/NCBI
23	Cui L, Xu S, Song Z, Zhao G, Liu X and Song Y: Pituitary tumor transforming gene: A novel therapeutic target for glioma treatment. Acta Biochim Biophys Sin (Shanghai). 47:414–421. 2015. View Article : Google Scholar : PubMed/NCBI
24	Jiang T, Qiao M, Zhou F, Ren S, Su C and Zhou C: Effect of combined therapy inhibiting EGFR and VEGFR pathways in non-small-cell lung cancer on progression-free and overall survival. Clin Lung Cancer. Dec 29–2016.(Epub ahead of print).
25	Yu X, Li W, Deng Q, You S, Liu H, Peng S, Liu X, Lu J, Luo X, Yang L, et al: Neoalbaconol inhibits angiogenesis and tumor growth by suppressing EGFR-mediated VEGF production. Mol Carcinog. 56:1414–1426. 2017. View Article : Google Scholar : PubMed/NCBI