Dados do Resumo

Título

Role of LGR5 Receptor in Prognosis and Biology of Soft Tissue Sarcomas

Introdução

LGR5 is a G protein-coupled receptor, an important class of membrane proteins responsible for transmitting extracellular signals. It acts as a negative modulator of the Wnt/β-catenin signaling pathway, implicating it with a putative role in biology of sarcomas.

Objetivo

Considering previous findings pointing to a modulation of Lgr5 upon cell migration and self-renewal, this project aims to generate Lgr5-deficient sarcoma cells in order to investigate its role in sarcoma biology.

Métodos

We employed the fibrosarcoma HT-1080 cell line to create LGR5-deficient cells using CRISPR technology. Specifically, the PX330 vector was digested and annealed with selected guides targeting the LGR5 gene. The resulting plasmids were transduced into competent bacteria, and colonies were isolated. The plasmids were then purified and sequenced. HT-1080 cells will be transfected with these plasmids, and flow cytometry will be used for analysis. In vitro assays will be conducted to assess migration, self-renewal, proliferation, and sensitivity to doxorubicin.

Resultados

We are currently generating HT-1080 LGR5-deficient clones. The plasmid carrying the LGR5 guides has been successfully produced. We are now analyzing plasmid sequencing data to proceed with cell transfection. The next steps involve characterizing the transfected cells to identify LGR5-deficient clones, followed by assessing their proliferation, migration, self-renewal, sensitivity to doxorubicin and in vivo tumor growth activity.

Conclusões

Our hypothesis is that Lg5 may influence the behavior of soft tissue sarcomas and could represent a potential therapeutic target for these tumors.

Palavras Chave

Soft Tissue Sarcomas; LGR5