Selection of highly malignant tumour cells using reconstituted basement membrane matrix

Area of Science:

• Biochemistry
• Cell Biology
• Oncology

Background:

• The basement membrane is a critical barrier in cancer metastasis.
• Identifying cancer cell subpopulations capable of traversing the basement membrane is essential for understanding metastasis.

Purpose of the Study:

• To develop a novel technique for coating filters with basement membrane matrix components.
• To utilize this technique for selecting and isolating cancer cell clones with enhanced metastatic potential.
• To assess the metastatic capacity of selected clones in vivo.

Main Methods:

• Extraction of proteoglycan and laminin from EHS tumor.
• Solubilization of collagen type IV.
• Formation of a basement membrane gel on Nuclepore filters using a flat-bed dialysis apparatus.
• Selection of B16 melanoma cells penetrating the coated filters.
• Assay of spontaneous metastasis in C57Bl/6 mice after intracutaneous injection.

Main Results:

• A reproducible method for coating filters with basement membrane components was established.
• Ten clones with significantly increased metastatic rates to lungs and lymph nodes were isolated.
• Selected cells demonstrated the ability to reach both accessible and less accessible metastatic sites.
• No specific preference for hematogenous or lymphatic routes was observed, indicating a general increase in metastatic ability.

Conclusions:

• The developed method is a valuable tool for isolating cancer cell subpopulations with high metastatic potential.
• The ability to traverse the basement membrane correlates with a general increase in metastatic capacity.
• This technique can be used to assess the metastatic potential of new tumor samples.