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Published on: September 13, 2022
Selection of internalizing RNA aptamers into human breast cancer cells derived from primary sites
Pricila da Silva Cunha1, Marcelo Coutinho de Miranda1, Mariane Izabella Abreu de Melo1
1Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Abstract:
Breast cancer is the most common cancer in women. Although chemotherapy is still broadly used in its treatment, adverse effects remain a challenge. In this scenario, aptamers emerge as a promising alternative for theranostic applications. Studies using breast cancer cell lines provide useful information in laboratory and preclinical investigations, most of which use cell lines established from metastatic sites. However, these cell lines correspond to cell populations of the late stage of tumor progression. On the other hand, studies using breast cancer cells established from primary sites make it possible to search for new theranostic approaches in the early stages of the disease. Therefore, this work aimed to select RNA aptamers internalized by MGSO-3 cells, a human breast cancer cell line, derived from a primary site previously established in our laboratory. Using the Cell-Internalization SELEX method, we have selected two candidate aptamers (ApBC1 and ApBC2). We evaluated their internalization efficiencies, specificities, cellular localization by Reverse Transcription-qPCR (RT-qPCR) and confocal microscopy assays. The results suggest that both aptamers were efficiently internalized by human breast cancer cells, MACL-1, MDA-MB-231, and especially by MGSO-3 cells. Furthermore, both aptamers could effectively distinguish human breast cancer cells derived from normal human mammary cell (MCF 10A) and prostate cancer cell (PC3) lines. Therefore, ApBC1 and ApBC2 could be promising candidate molecules for theranostic applications, even in the early stages of tumor progression.
Insights
Researchers identified two RNA aptamers, ApBC1 and ApBC2, that are internalized by early-stage breast cancer cells. These aptamers show promise for developing new theranostic tools for breast cancer treatment.
Area of Science:
- Biotechnology
- Molecular Biology
- Oncology
Background:
- Breast cancer is a leading cause of cancer in women, with chemotherapy often causing adverse effects.
- Current research often uses cell lines from metastatic sites, representing late-stage disease.
- Theranostic applications for early-stage breast cancer require cell lines derived from primary tumors.
Purpose of the Study:
- To select and characterize RNA aptamers that are internalized by MGSO-3 cells, a human breast cancer cell line derived from a primary site.
- To evaluate the potential of these aptamers for theranostic applications in early-stage breast cancer.
Main Methods:
- Cell-Internalization SELEX (Systematic Evolution of Ligands by Exponential Enrichment) was used to select aptamers.
- Internalization efficiency, specificity, and cellular localization were assessed using RT-qPCR and confocal microscopy.
- Candidate aptamers were tested against various human cancer and normal cell lines.
Main Results:
- Two RNA aptamers, ApBC1 and ApBC2, were successfully selected.
- Both aptamers demonstrated efficient internalization by multiple human breast cancer cell lines, particularly MGSO-3 cells.
- ApBC1 and ApBC2 showed high specificity, distinguishing breast cancer cells from normal mammary and prostate cancer cells.
Conclusions:
- ApBC1 and ApBC2 are promising candidate molecules for theranostic applications in breast cancer.
- These aptamers show potential for targeting early-stage breast cancer due to their internalization by primary site-derived cells.
- The findings support the development of novel aptamer-based diagnostic and therapeutic strategies for breast cancer.
