Tumor Biology

, Volume 37, Issue 3, pp 3797–3805 | Cite as

The vascular delta-like ligand-4 (DLL4)-Notch4 signaling correlates with angiogenesis in primary glioblastoma: an immunohistochemical study

  • Jin-feng Zhang
  • Yao Chen
  • Xian-xin Qiu
  • Wen-long Tang
  • Jian-dong Zhang
  • Jian-huang Huang
  • Guo-shi Lin
  • Xing-fu Wang
  • Zhi-xiong Lin
Original Article
First Online:
Received:
Accepted:

Abstract

Delta-like ligand-4 (DLL4)-Notch signaling is known to play a pivotal role in the regulation of tumor angiogenesis. We had previously found that DLL4 was overexpressed, while Notch1 receptor, which binds to DLL4 during angiogenesis, was absent in the majority of human primary glioblastomas. Thus, DLL4-Notch signaling pathway in the regulation of tumor angiogenesis in primary glioblastoma remains unknown. Tumor tissues from 70 patients with primary glioblastoma were analyzed by immunohistochemistry for expression of components of DLL4-Notch signaling, vascular endothelial growth factor (VEGF), and microvessel density (MVD). Immunohistochemistry results showed that the positive staining of DLL4 and Notch4 was primarily distributed in tumor vascular endothelial cells but rarely detected in tumor cells. However, VEGF, hairy/enhancer of split-1 (HES1; a target gene of Notch signaling), and Notch1–3 expression was seen in both tumor vascular endothelial cells and tumor cells. Univariate analysis showed that the expression levels of VEGF and DLL4, HES1, and Notch4 in tumor endothelial cells were significantly associated with MVD in primary glioblastoma (P < 0.001). Binary logistic regression analysis showed that high expression levels of DLL4, HES1, and Notch4 in tumor endothelial cells were associated with a decrease of MVD in primary glioblastoma, while MVD increased with elevated VEGF expression in contrast. In addition, DLL4, Notch4, and HES1 expression were positively correlated in tumor vascular endothelial cells (P < 0.05). We conclude that the vascular DLL4-Notch4 signaling and VEGF signaling complementing each other plays an important role in the progression of tumor angiogenesis in primary glioblastoma.

Graphical abstract

A, positive staining of DLL4 in human kidney; B, positive staining of VEGF in human breast cancer; C, positive staining of CD34 in human lung cancer; D, positive staining of HES1 in human breast cancer; E-H, positive staining of Notch1-4: E-F in human lung cancer; G-H in human kidney

Keywords

Primary glioblastoma DLL4-Notch signaling Vascular endothelial growth factor Microvessel density Angiogenesis 

Abbreviations

MVD

Microvessel density

VEGF

Vascular endothelial growth factor

DLL4

Delta-like ligand-4

NICD

Notch intracellular domain

WHO

World Health Organization

PBS

Phosphate buffer solution

DAB

Diaminobenzidine

EC

Endothelial cells

TC

Tumor cell

GSC

Glioma stem cell

PI3k

Phosphatidylinositol 3-kinase

EGFR

Epidermal growth factor receptor

MMP-9

Matrix metalloprotein-9

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Notes

Acknowledgments

This study was supported by the National Natural Science Foundation of China (No. 21435002). The authors would like to thank the Department of Pathology, First Affiliated Hospital, Fujian Medical University, for their help in immunohistochemistry design and the Public Health School, Fujian Medical University, Fujian, China, for assistance in data processing and statistical analysis.

Authors’ contributions

JFZ and YC analyzed the data and drafted the manuscript. YC, JFZ, and XXQ performed the experiments. XFW and JDZ provided material support. WLT, GSL, and JHH contributed to data collection and helped in the analysis. ZXL conceived the study and participated in its design and coordination. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Jin-feng Zhang
    • 1
  • Yao Chen
    • 2
  • Xian-xin Qiu
    • 3
  • Wen-long Tang
    • 2
  • Jian-dong Zhang
    • 2
  • Jian-huang Huang
    • 2
  • Guo-shi Lin
    • 2
  • Xing-fu Wang
    • 4
  • Zhi-xiong Lin
    • 1
    • 5
  1. 1.The First Clinical Medical CollegeFujian Medical UniversityFuzhouChina
  2. 2.Department of Neurosurgery, The First Affiliated HospitalFujian Medical UniversityFuzhouChina
  3. 3.Department of Neurosurgery, The Children’s Hospital, School of MedicineZhejiang UniversityHangzhouChina
  4. 4.Department of Pathology, The First Affiliated HospitalFujian Medical UniversityFuzhouChina
  5. 5.Department of Neurosurgery, Beijing Sanbo Brain HospitalCapital Medical UniversityBeijingChina

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