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Preparation of Mitochondria from Ovarian Cancer Tissues and Control Ovarian Tissues for Quantitative Proteomics Analysis
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Spatial-Resolution Cell Type Proteome Profiling of Cancer Tissue by Fully Integrated Proteomics Technology
Ruilian Xu1, Jun Tang1,2, Quantong Deng1
1Shenzhen People's Hospital , The Second Clinical Medical College of Jinan University , Shenzhen 518020 , China.
Analytical Chemistry
|April 12, 2018
Summary
This study introduces LCM-SISPROT, a new method for high-sensitivity cell type proteome profiling from minimal tumor tissue. It enables precise dissection and analysis of specific cell types within the tumor microenvironment.
Area of Science:
- Proteomics
- Molecular Biology
- Cancer Research
Background:
- Understanding the tumor microenvironment requires cell type-specific proteome profiling.
- Current methods demand large sample amounts, limiting clinical tumor specimen analysis.
Purpose of the Study:
- To develop a high-sensitivity proteome profiling method for minute clinical tumor samples.
- To enable precise cell type dissection and analysis within the tumor microenvironment.
Main Methods:
- Combined laser capture microdissection (LCM) with integrated proteomics sample preparation (SISPROT).
- Developed a noncontact, fully integrated system to minimize sample loss and contamination.
- Utilized a two-stage spintip device for selective removal of H&E staining dyes.
Main Results:
- Achieved high-sensitivity proteome profiling, identifying >500 proteins from 0.1 mm² of colon cancer tissue.
- Successfully performed cell type proteome profiling of cancer cells, enterocytes, lymphocytes, and smooth muscle cells from a single tumor.
- Identified thousands of protein groups from minute tissue sections (5 mm², 10 μm thickness).
- Demonstrated spatially resolved proteome distribution profiles with micrometer precision.
Conclusions:
- LCM-SISPROT enables precise, high-sensitivity cell type proteome profiling using minimal clinical samples.
- The technology is promising for detailed spatial analysis of the tumor microenvironment.
- Facilitates understanding of tumor heterogeneity and cellular interactions.
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