Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and <em>Ex vivo</em> Fibroblasts

Marc D. Bullock; Max Mellone; Karen M. Pickard; Abdulkadir Emre Sayan; Richard Mitter; John N. Primrose; Graham K. Packham; Gareth Thomas; Alexander H. Mirnezami

doi:10.3791/51475

Moleküler Kolorektal Kanser Hücrelerinin bir 3-Boyutlu Modeli İnvaziv tümör mikro Profili ve ...

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JoVE Journal Medicine

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

Moleküler Kolorektal Kanser Hücrelerinin bir 3-Boyutlu Modeli İnvaziv tümör mikro Profili ve Ex vivo Fibroblastlar

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10:33 min

April 29, 2014

DOI: 10.3791/51475-v

Marc D. Bullock^1,2, Max Mellone¹, Karen M. Pickard¹, Abdulkadir Emre Sayan¹, Richard Mitter³, John N. Primrose², Graham K. Packham¹, Gareth Thomas¹, Alexander H. Mirnezami^1,2

¹Cancer Sciences Unit,University of Southampton School of Medicine, ²University Surgical Unit,University of Southampton School of Medicine, ³Bioinformatics Unit,London Research Institute, Cancer Research UK

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Overview

This study presents a novel approach to characterize the biology at the interface between malignant epithelial cells and cancer-associated stromal cells in colorectal cancer models. By utilizing laser microdissection, researchers can dissect and analyze the distinct cellular environments within tumor microenvironments.

Key Study Components

Area of Science

Neuroscience
Oncology
Cell Biology

Background

Colorectal cancer involves complex interactions between tumor cells and the surrounding stroma.
Understanding these interactions is crucial for developing targeted therapies.
Laser microdissection allows for precise isolation of specific cell types within tumors.
This methodology can enhance molecular profiling efforts.

Purpose of Study

To characterize the biology at the invasive front of colorectal tumors.
To differentiate between invading and non-invading tumor cells.
To analyze the role of cancer-associated stromal cells in tumor progression.

Methods Used

Establishment of primary fibroblast cultures from colonic implants.
Construction of organotypic co-culture models with fibroblasts and colorectal cancer cells.
Fixation and sectioning of organotypic models for analysis.
Laser microdissection to separate tumor and stromal cells.

Main Results

Molecular profiling reveals distinct biological characteristics of tumor and stromal cells.
Identified differences in gene expression between invading and non-invading cells.
Insights into the tumor microenvironment's role in cancer progression.
Potential for developing targeted therapeutic strategies based on these findings.

Conclusions

This approach provides a manipulatable model for studying colorectal cancer biology.
Laser microdissection is a valuable tool for isolating specific cell populations.
Understanding the tumor-stroma interface can inform future cancer therapies.

Frequently Asked Questions

What is laser microdissection?

Laser microdissection is a technique used to isolate specific cells from a tissue sample using a laser.

Why is it important to study the tumor microenvironment?

The tumor microenvironment plays a critical role in cancer progression and response to therapy.

How do organotypic co-culture models work?

These models simulate the interactions between different cell types in a controlled environment, mimicking in vivo conditions.

What are the potential applications of this research?

Findings could lead to new therapeutic strategies targeting the tumor-stroma interactions in colorectal cancer.

What are cancer-associated stromal cells?

These are non-cancerous cells in the tumor microenvironment that can influence tumor growth and behavior.

What is the significance of profiling invading vs. non-invading cells?

Profiling these cells helps to understand the mechanisms of cancer invasion and metastasis.

Sentetik, doku mühendisliği kolorektal kanser modelleri, lazer microdissected stroma hücreleri ve moleküler profil invaziv ön ve kanserle ilişkili stromal hücre tümör hücreleri arasındaki ara yüzeyde ayırt edici karakterize biyoloji ve incelemek için bir yeni ve manuel olarak kullanılabilen bir yaklaşımı temsil etmektedir.

Bu prosedürün genel amacı, invaziv tümör cephesindeki malign epitel hücreleri ile kanserle ilişkili stromal hücreler arasındaki arayüzdeki ayırt edici biyolojiyi karakterize etmektir. Bu, ilk olarak kolon implantlarından primer fibroblast kültürlerinin oluşturulmasıyla gerçekleştirilir. İkinci adım, primer kolon fibroblastları ve kültürlenmiş kolorektal kanser hücrelerini kullanarak organotipik ko-kültür modelleri oluşturmaktır.

Daha sonra, organotipik modeller formin içinde sabitlenir, membrana monte slaytlar üzerine kesitlenir ve krestal menekşe ile boyanır. Son adım, invaziv tümör hücrelerini invaziv olmayan tümör hücrelerinden ve kanserle ilişkili stromal hücrelerden ayırmak için lazer mikrodiseksiyondur. Sonuç olarak, moleküler profilleme teknikleri, çevredeki tümör mikroçevresindeki istilacı ve invaziv olmayan tümör hücrelerinin ve stromal hücrelerin biyolojisini karakterize etmek için kullanılabilir.

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