1/3. Double mutation and gene copy number of EGFR in gefitinib refractory non-small-cell lung cancer.Mutations of the epidermal growth factor receptor (EGFR) gene have been reported in non-small-cell lung cancer (NSCLC), especially in patients with adenocarcinoma and never smokers. Some common somatic mutations in EGFR, including deletion mutations in exon 19 and leucine-to-arginine substitution at amino acid position 858 (L858R) in exon 21, have been examined for their ability to predict sensitivity to gefitinib or erlotinib, which are selective EGFR tyrosine kinase inhibitors (EGFR-TKIs). On the other hand, reports have shown that the threonine-to-methionine substitution at amino acid position 790 (T790M) in exon 20 is related to gefitinib resistance. Some studies have indicated that high copy numbers of the EGFR gene may be a more effective molecular predictor to responsiveness and prolonged survival in patients treated with EGFR-TKIs. Here, we describe two NSCLC patients with the L858R mutation who did not respond to gefitinib. Case 1 harbored both the T790M and L858R mutations, and fluorescence in situ hybridization showed EGFR gene amplification. Case 2 harbored both the L858R and aspartic acid-to-tyrosine substitution at amino acid position 761 in exon 19 of EGFR mutations and had a high polysomy status for EGFR. In these two cases, tumors showed resistance to gefitinib treatment despite the presence of EGFR L858R mutation and increased copy number. Our findings encourage further molecular analysis to elucidate the relationship between the EGFR status, including mutations and amplifications, and the responsiveness of NSCLC to gefitinib.- - - - - - - - - - ranking = 1keywords = hybridization (Clic here for more details about this article) |
2/3. Analysis of abnormal expression of g-csf gene in a novel tumor cell line (KHC 287) elaborating G-CSF, IL-1 and IL-6 with co-amplification of c-myc and c-ki-ras.We established a human carcinoma cell line (KHC 287) from a patient with large-cell-typing lung carcinoma associated with marked leukocytosis. The culture supernatant of KHC 287 cells promoted granulocytic colony formation of human bone-marrow cells in semi-solid culture. Colony formation was almost completely suppressed by treatment of the supernatant with a monoclonal anti-granulocyte colony-stimulating factor (G-CSF) antibody. Not only G-CSF but also interleukin-1 alpha (IL-I alpha), IL-I beta and IL-6 were detected in the culture supernatant by an ELISA method. Northern blot analysis of KHC 287 cells revealed distinct expression of these cytokine genes. Southern blot hybridization of KHC 287 dna showed 20- and 40-fold co-amplification of c-myc and c-ki-ras, respectively. The chloramphenicol acetyl transferase (CAT) activity was distinctly enhanced in the KHC 287 cells which were transfected with the 360 bp upstream region of G-CSF gene inserted into pSV00CAT, but not in non-G-CSF-producing tumor cell lines. These results suggest that overproduction of the transactivating factor(s) which binds to the 360 bp of the G-CSF upstream region is responsible for the abnormal expression of G-CSF gene in KHC 287 cell line.- - - - - - - - - - ranking = 1keywords = hybridization (Clic here for more details about this article) |
3/3. Characterization of 9q;15q whole-arm translocation derivatives in non-small cell lung carcinomas by fluorescence in situ hybridization.We report derivative chromosomes, originally interpreted as 9q;15q whole-arm translocations, in tumor cells from two patients with non-small cell lung cancer (NSCLC). One of the tumors was diagnosed as an adenocarcinoma and the other as an adenosquamous carcinoma. In each case, there was no normal chromosome 9. Because of the pericentromeric location of the breakpoints, classical cytogenetic banding techniques did not permit determination of the centromeric origin of these derivative chromosomes. fluorescence in situ hybridization (FISH) with satellite (alpha, beta, classical), ribosomal dna, alpha-interferon (alpha-IFN), and whole chromosome painting probes indicated that the 9;15 rearrangement is dicentric in both tumors. In one of these cases, the derivative chromosome is interpreted as a dic(9;15) (p11;p11.2); the other case has a more complicated rearrangement involving reorientation of pericentromeric sequences. A 9q;15q whole-arm derivative chromosome was reported previously in another lung adenocarcinoma, suggesting that this abnormality may represent a recurrent change in lung carcinomas, particularly those displaying adenomatous features.- - - - - - - - - - ranking = 5keywords = hybridization (Clic here for more details about this article) |