Preparation Of Neovascular Tissues from Human Glioma Tissues for Quantitative Proteomics Analysis of Tumor Angiogenesis

Xiaoxia Feng; Zheng Fang; Xiaohan Zhan; Tianyao Guo; Yanjun Zhang; Zhuoran Yin; Yuping Zhang; Na Li; Xianquan Zhan

doi:10.3791/69547

A subscription to JoVE is required to view this content. Sign in or start your free trial.

Method Article

Preparation Of Neovascular Tissues from Human Glioma Tissues for Quantitative Proteomics Analysis of Tumor Angiogenesis

547 views

⸱

DOI:

10.3791/69547

⸱

March 20th, 2026

Xiaoxia Feng¹^,² , Zheng Fang¹^,² , Xiaohan Zhan³ , Tianyao Guo⁴^,⁵ , Yanjun Zhang¹^,² , Zhuoran Yin¹^,² , Yuping Zhang⁶ , Na Li¹ , Xianquan Zhan¹^,²

¹Shandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University & Shandong Academy of Medical Science, ²Shandong Provincial Key Medical and Health Laboratory of Ovarian Cancer Multiomics, & Jinan Key Laboratory of Cancer Multiomics, Shandong First Medical University & Shandong Academy of Medical Science, ³School of Medicine, Wayne State University, ⁴Department of Pathology, The Second Xiangya Hospital, Central South University, ⁵Hunan Clinical Medical Research Center for Cancer Pathogenic Genes Testing and Diagnosis,, ⁶Department of The 1st General Surgery, The Third Xiangya Hospital, Central South University

Summary

This protocol successfully prepared sufficient high-quality neovascular tissue from glioma samples using laser-capture microdissection for quantitative proteomics analysis of tumor angiogenesis, overcoming the limitations of studies that traditionally focus mainly on angiogenic factors and providing a large-scale proteomics profile of glioma angiogenesis.

Abstract

Glioma represents primary malignant tumors of the central nervous system with significant incidence and mortality. Tumor angiogenesis is the hallmark of glioma pathophysiology and is an important event in the tumor environment. The aberrant structure of tumor neovascularization serves dual roles: a nutrient supply source and a metastatic portal enabling tumor cell invasion into the bloodstream. Anti-angiogenesis therapy is a hot topic in the field of cancer. Most of the previous studies focus on the activating factors and inhibitory factors of tumor angiogenesis; however, this only clarifies tumor angiogenesis in a very limited range. Abnormal protein expression or post-translational modifications contribute to the abnormal morphology of blood vessels in glioma. This protocol used laser-capture microdissection (LCM) to isolate and purify an adequate amount of neovascular tissues from glioma tissues, which was used for isotope-labelled quantitative proteomics analysis to identify differentially expressed proteins in glioma neovascular tissues compared to controls. These data provide a crucial scientific basis for elucidating the pathological mechanisms of tumor angiogenesis, discovering anti-angiogenic therapeutic targets, and constructing tumor angiogenesis-based biomarkers.

Introduction

Gliomas are highly heterogeneous central nervous system(CNS) tumors originating from glial cells that provide structural and metabolic support to neurons¹, and are histologically classified into subtypes: astrocytomas, oligodendrogliomas, and ependymomas². According to the WHO grading system, glioma grades I and II are the low-grade gliomas (LGG), typically characterized by slow growth rates and potential long-term stability, often amenable to surgical resection based on anatomical location. In contrast, grades III and IV are the high-grade gliomas (HGG), predominantly undifferentiated and malignant with highly invasive ....