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Quantitative Immunohistochemistry of the Cellular Microenvironment in Patient Glioblastoma Resections
Published on: July 31, 2017
Quantitation of macropinocytosis in glioblastoma based on high-content analysis
Bo Wang1, Xuan Yao1, Qiang Dong2
1Key Lab of Neurology of Gansu Province, the Second Hospital of Lanzhou University, Lanzhou 730030, Gansu, China; The Department of Neurosurgery, the Second Hospital of Lanzhou University, Lanzhou 730030, Gansu, China; The Second Medical College of Lanzhou University, Lanzhou,730030, Gansu, China.
Background:
Macropinocytosis is a pathway utilized for the internalization of extracellular fluid, albumin and dissolved molecules. Assessing macropinocytosis has been challenging in the past because the combination of manual acquisition and visual evaluation of images is laborious, making this type of assessment difficult for high-throughput applications. Therefore, there is a need to develop sensitive and specific macropinocytosis evaluation methods.
Methods:
This paper proposed a quantitative and time-saving method for macropinocytosis detection based on high-content analysis (HCA). Additionally, cell proliferation was evaluated using CCK8 test.
Results:
The term "macropinosome index" was defined to estimate macropinocytosis and allow comparisons between different cell lines and treatments. Furthermore, we demonstrated that macropinocytosis can promote glioblastoma (GBM) cell survival under L-glutamine (L-Gln)-deficient conditions that resemble the tumour microenvironment.
Conclusions:
HCA represents a novel, nonsubjective and high-throughput assay for macropinocytosis assessment. In addition, L-Gln deprivation increased the macropinosome index in GBM cells, suggesting that this process may be used to design GBM therapies.
Availability Of Data And Materials:
The datasets supporting the conclusions of this article are included within the article and its supplementary materials.
Insights
A new high-content analysis method quantifies macropinocytosis, a cellular process. This technique aids in understanding glioblastoma survival mechanisms and developing novel therapies.
Area of Science:
- Cell Biology
- Biotechnology
Background:
- Macropinocytosis is crucial for cellular uptake but difficult to assess at scale.
- Existing methods are laborious and unsuitable for high-throughput screening.
- Development of sensitive and specific assays for macropinocytosis is needed.
Purpose of the Study:
- To introduce a quantitative, time-saving method for macropinocytosis detection using high-content analysis (HCA).
- To evaluate the role of macropinocytosis in glioblastoma (GBM) cell survival under nutrient-deficient conditions.
Main Methods:
- High-content analysis (HCA) was employed for quantitative macropinocytosis detection.
- A 'macropinosome index' was defined for standardized assessment.
- Cell proliferation was assessed using the CCK8 assay.
Main Results:
- The HCA method provides a novel, objective, and high-throughput assay for macropinocytosis.
- Macropinocytosis was shown to enhance GBM cell survival in L-glutamine-deficient environments.
- An increased macropinosome index was observed in GBM cells under L-glutamine deprivation.
Conclusions:
- HCA offers a robust platform for macropinocytosis assessment.
- Macropinocytosis plays a role in GBM cell adaptation to the tumor microenvironment.
- Targeting macropinocytosis presents a potential therapeutic strategy for GBM.
