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Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells
Published on: May 20, 2015
ADAM9 silencing inhibits breast tumor cell invasion in vitro
Kelli Cristina Micocci1, Ana Carolina Baptista Moreno Martin, Cyntia de Freitas Montenegro
1Departamento de Ciências Fisiológicas, Rodovia Washington Luís, Km 235, CEP 13565-905, São Carlos, SP, Brazil.
Abstract:
ADAM9 (A Disintegrin And Metalloproteinase 9) is a member of the ADAM protein family which contains a disintegrin domain. This protein family plays key roles in many physiological processes, including fertilization, migration, and cell survival. The ADAM proteins have also been implicated in various diseases, including cancer. Specifically, ADAM9 has been suggested to be involved in metastasis. To address this question, we generated ADAM9 knockdown clones of MDA-MB-231 breast tumor cells using silencing RNAs that were tested for cell adhesion, proliferation, migration and invasion assays. In RNAi-mediated ADAM9 silenced MDA-MB-231 cells, the expression of ADAM9 was lower from the third to the sixth day after silencing and inhibited tumor cell invasion in matrigel by approximately 72% when compared to control cells, without affecting cell adhesion, proliferation or migration. In conclusion, the generation of MDA-MB-231 knockdown clones lacking ADAM9 expression inhibited tumor cell invasion in vitro, suggesting that ADAM9 is an important molecule in the processes of invasion and metastasis.
Insights
A Disintegrin And Metalloproteinase 9 (ADAM9) knockdown in breast cancer cells significantly inhibited tumor invasion by 72%. This suggests ADAM9 is crucial for cancer metastasis and invasion processes.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- A Disintegrin And Metalloproteinase 9 (ADAM9) is part of the ADAM protein family, known for roles in cell migration and survival.
- ADAM proteins are implicated in various diseases, including cancer, with ADAM9 specifically linked to metastasis.
Purpose of the Study:
- To investigate the role of ADAM9 in breast tumor metastasis.
- To determine if reducing ADAM9 expression affects cancer cell invasion, adhesion, proliferation, and migration.
Main Methods:
- Generated ADAM9 knockdown clones of MDA-MB-231 breast tumor cells using RNA interference (RNAi).
- Assessed cell adhesion, proliferation, migration, and invasion (in matrigel) in ADAM9-silenced cells compared to controls.
Main Results:
- ADAM9 expression was significantly reduced in knockdown cells from day 3 to day 6 post-silencing.
- Tumor cell invasion in matrigel was inhibited by approximately 72% in ADAM9-silenced cells.
- Cell adhesion, proliferation, and migration remained unaffected by ADAM9 knockdown.
Conclusions:
- Knockdown of ADAM9 in MDA-MB-231 cells effectively inhibits in vitro tumor cell invasion.
- ADAM9 is identified as a key molecule involved in the processes of cancer invasion and metastasis.
