Imatinib-drug-resistance KIT and PDGFRA wild type GIST cell strain and establishment method and application thereof

Abstract

The invention provides an Imatinib-drug-resistance KIT and PDGFRA wild type GIST cell strain which is named as GIST-FR and preserved in the China Center for Type Culture Collection with the preservation number being CCTC NO:C2017111. The invention further provides an establishment method of the cell strain and application of the cell strain as a cell model for discussing the human GIST drug resistance mechanism and researching relevant signal channels thereof. A tissue block culture method is adopted to obtain GIST primary cells, immortalization is carried out, and a GIST-F cell line is established; then Imatinib drug resistance induction is carried out on the GIST-F cell line, and the Imatinib-drug-resistance GIST cell strain GIST-FR is established. The GIST-FR cells are subjected to in-vitro passage for more than 40 generations, growth is slow compared with GIST-F cells, IC50 of Imatinib is remarkably increased, and the drug resistance index is 3.52 (P is less than 0.01); and a wholeexon group sequencing result shows that KIT and PDGFRA of the GIST-FR cells are negative. The GIST cell line is successfully built from human GIST primary tissue, and further induction is carried outto obtain the Imatinib-drug-resistance cells thereof, so that a foundation is laid for the GIST pathogenesis and drug resistance related research.

Description

technical field [0001] The present invention relates to a drug-resistant cell line and its construction method and application, in particular to an Imatinib-resistant KIT and PDGFRA wild-type GIST cell line and its construction method and application. Background technique [0002] GIST (gastrointestinal stromal tumor) is the most common mesenchymal tumor in the gastrointestinal tract, accounting for about 1% to 3% of malignant tumors in the gastrointestinal tract. Studies have found that about 85% of gastrointestinal stromal tumors (GIST) have KIT (65%) or PDGFRA (20%) gene mutations. Both KIT and PDGFRA belong to the transmembrane type III tyrosine receptor family. [0003] At present, research on the pathogenesis, pathological diagnosis, and clinical treatment of gastrointestinal stromal tumors (GIST) has made great progress. Tyrosine kinase inhibitors, mainly imatinib, etc. The application of molecular targeted drugs has greatly improved the clinical efficacy of gastroi...

AI Technical Summary

Problems solved by technology

[0005] In view of the above problems, the present invention provides an Imatinib-resistant KIT and PDGFRA wild-type GIST cell line and its construction method and application, so as to overcome the current domestic imatinib-resistant KIT and PDGFRA wild-type GIST resistance. Drug-resistant cell lines make it difficult to study the pathogenesis and drug resistance mechanism of GIST, thus laying the foundation for research on GIST drug resistance and increasing the influence of the country in international medicine

Images