Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional (3D) Model

Donna Cvetkovi&#263;; Cameron Glenn-Franklin Goertzen; Moshmi Bhattacharya

doi:10.3791/51341

Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional (3D) Model

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Medicine

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JoVE Journal Medicine

Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional (3D) Model

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08:08 min

June 11, 2014

DOI: 10.3791/51341-v

Donna Cvetković*¹, Cameron Glenn-Franklin Goertzen*¹, Moshmi Bhattacharya^1,2,3

¹Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry,University of Western Ontario, ²Department of Oncology, Schulich School of Medicine and Dentistry,University of Western Ontario, ³Lawson Health Research Institute

Overview

This article provides methodologies for three-dimensional (3D) assays to quantify breast cancer cell invasion. The procedures include setting up assays, quantification, and data analysis, along with examining membrane integrity loss during cell invasion.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Cancer Research

Background

Invasive ability of cancer cells is critical for understanding metastasis.
3D assays better mimic the in vivo environment compared to traditional methods.
Fluorescent labeling allows for detailed protein localization studies.
Light microscopy is used to observe cellular invasion over time.

Purpose of Study

To evaluate the invasive ability of breast cancer cells.
To analyze changes in protein expression during invasion.
To improve methodologies for studying cancer cell behavior.

Methods Used

Culturing cancer cells in a basement membrane matrix.
Observing cellular invasion via light microscopy for five or more days.
Labeling cells with fluorescent antibodies for protein localization.
Analyzing invasion rates and protein expression changes using immunofluorescent microscopy.

Main Results

The 3D invasion assay effectively mimics the in vivo environment.
Significant changes in protein expression were observed during invasion.
The method provides a reliable quantification of cancer cell invasion.
Results indicate enhanced understanding of cancer cell behavior.

Conclusions

The 3D major gel invasion assay is superior to traditional techniques.
This method can lead to better insights into cancer metastasis.
Future studies can build on these methodologies for further research.

Frequently Asked Questions

What is the main advantage of the 3D invasion assay?

The 3D invasion assay better mimics the in vivo environment compared to traditional methods like the Transwell chamber.

How long do you observe the cellular invasion?

Cellular invasion is observed for five or more days using light microscopy.

What technique is used for protein localization?

Fluorescent antibodies are used for the localization of proteins of interest.

What is the purpose of labeling cells with antibodies?

Labeling allows for the visualization and analysis of protein expression changes during invasion.

How does this method contribute to cancer research?

It provides a more accurate representation of cancer cell behavior and invasion, aiding in the understanding of metastasis.

Can this method be applied to other types of cancer?

Yes, the 3D invasion assay can be adapted for various cancer types to study their invasive properties.

This article provides detailed methodologies for the use of three-dimensional (3D) assays to quantify breast cancer cell invasion. Specifically, we discuss the procedures required to set up such assays, quantification, and data analysis, as well as methods to examine the loss of membrane integrity that occurs when cells invade.

The overall goal of the following procedure is to evaluate the invasive ability of cancer cells using a three dimensional invasion assay. This is accomplished by first culturing cancer cells in a basement membrane matrix. The cellular invasion of the matrix is then observed for five or more days via light microscopy.

In the final step, the cells are labeled with fluorescent antibodies for localization of the proteins of interest. Ultimately, the rate of cancer cell invasion and the resulting changes in protein expression can be analyzed by immunofluorescent microscopy. The main advantage of this technique over existing techniques such as the Transwell chamber invasion assay, is that the 3D major gel invasion assay better mimics the in vivo environment that cancer solves growing.

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