Method of diagnosing, classifying and treating endometrial cancer and precancer
A technology for endometrial cancer and endometrium, which is applied in the field of diagnosis, classification and treatment of endometrial cancer and early cancer, and can solve the problems of poor survival rate, unproven adjuvant therapy, difficult to cure tumor chemotherapy, etc.
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Embodiment 1
[0141] Detection of activating FGFR2 mutations in endometrial cancer
[0142] Our findings show that activation and overexpression of FGFR2 plays a role in endometrial tumorigenesis. Exon 8 is 3 nucleotides longer than exon 9, so the FGFR2b isoform is one codon longer than the FGFR2c isoform. Specificity of signaling is also provided by tissue-specific expression of receptors, ligands and heparin sulphate proteoglycans (Allen et al., 2001; Fiore, 2001). Due to the difference in length of the FGFR2 "b" and "c" isoforms, all mutations are numbered relative to the epithelially expressed FGFR2b isoform (SEQ ID NO: 2; NP_075259.2). For those occurring downstream of exon 8, here we provide the corresponding mutations numbered relative to the FGFR2c isoform (SEQ ID NO: 3; NP_000132.1) in brackets and in Table 2. The N550K (N549K) variant identified in two endometrial cell lines may result in an association with FGFR2 and FGFR3 in patients with Crouzon syndrome (Kan et al., 2002) ...
Embodiment 2
[0163] Treatment of endometrial cancer by inhibiting FGFR2
[0164] Materials and methods
[0165] Sequencing analysis
[0166] Mutation analysis was performed as previously described (8). PCR primer sequences were M13 tailed and sequencing was performed in both directions. Primer sequences were obtained by request from the authors.
[0167] Cell Culture and Reagents
[0168]The human endometrial MFE296 cell line was purchased from the European Collection of Cell Cultures (Salisbury, Wiltshire, UK). Human endometrial cell lines AN3CA, HEC1A, Ishikawa, RL952 and KLE were provided by Dr. Paul Goodfellow (Washington University, St. Louis, MO). MFE296 cells were grown in MEM supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine and penicillin-streptomycin. AN3CA cells were cultured in DMEM supplemented with 10% FBS, non-essential amino acids, 2 mM L-glutamine and penicillin-streptomycin. HEC1A cells were grown in 50% DMEM and 50% RPMI 1640 supplemented with 10%...
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