- MeSH
- bronchogenní karcinom * patologie MeSH
- diagnostické techniky molekulární metody MeSH
- geny erbB-1 genetika imunologie MeSH
- inhibitory proteinkinas terapeutické užití MeSH
- lidé MeSH
- mutační analýza DNA MeSH
- nádorové biomarkery analýza MeSH
- polymerázová řetězová reakce metody MeSH
- sekvenční analýza DNA MeSH
- Check Tag
- lidé MeSH
- MeSH
- alely * MeSH
- genetická predispozice k nemoci * MeSH
- genetické asociační studie * MeSH
- genotyp MeSH
- leukemie diagnóza etiologie terapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikrosatelitní repetice MeSH
- nádorové biomarkery * MeSH
- polymorfismus genetický MeSH
- transplantace hematopoetických kmenových buněk MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- dopisy MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
We herein present a rare case of an EML4-ALK positive patient. A 61-year-old man was diagnosed with locoregional non-small cell lung cancer (NSCLC). No EGFR mutations were detected, and therefore the ALK rearrangement was evaluated using immunohistochemistry (IHC), fluorescence in situ hybridization (FISH) and the reverse transcription PCR (RT-PCR) method for EML4-ALK. All methods showed a positive result and, therefore, the patient was treated with crizotinib with a good therapeutic response. However, a detailed RT-PCR analysis and sequencing revealed an unexpected 138 bp insertion of attractin-like 1 (ATRNL1) gene into the EML4-ALK fusion gene. In our case, the positive therapeutic response suggests that ATRNL1 insertion does not affect EML4-ALK's sensitivity to crizotinib. This case shows great EML4-ALK heterogeneity and also that basic detection methods (IHC, FISH) cannot fully specify ALK rearrangement but in many cases a full specification seems to be important for an effective TKI indication, and sequencing ALK variants might contribute to optimized patient selection.
- MeSH
- fúzní onkogenní proteiny chemie genetika MeSH
- inhibitory proteinkinas terapeutické užití MeSH
- inzerční mutageneze * MeSH
- lidé středního věku MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- nádory plic diagnóza farmakoterapie genetika MeSH
- nemalobuněčný karcinom plic diagnóza farmakoterapie genetika MeSH
- pyrazoly terapeutické užití MeSH
- pyridiny terapeutické užití MeSH
- rentgendiagnostika hrudníku MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- staging nádorů MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
OBJECTIVES: Lung cancer with the ALK rearrangement constitutes only a small fraction of patients with non-small cell lung cancer (NSCLC). However, in the era of molecular-targeted therapy, efficient patient selection is crucial for successful treatment. In this context, an effective method for EML4-ALK detection is necessary. We developed a new highly sensitive variant specific TaqMan based real time PCR assay applicable to RNA from formalin-fixed paraffin-embedded tissue (FFPE). MATERIALS AND METHODS: This assay was used to analyze the EML4-ALK gene in 96 non-selected NSCLC specimens and compared with two other methods (end-point PCR and break-apart FISH). RESULTS: EML4-ALK was detected in 33/96 (34%) specimens using variant specific real time PCR, whereas in only 23/96 (24%) using end-point PCR. All real time PCR positive samples were confirmed with direct sequencing. A total of 46 specimens were subsequently analyzed by all three detection methods. Using variant specific real time PCR we identified EML4-ALK transcript in 17/46 (37%) specimens, using end-point PCR in 13/46 (28%) specimens and positive ALK rearrangement by FISH was detected in 8/46 (17.4%) specimens. Moreover, using variant specific real time PCR, 5 specimens showed more than one EML4-ALK variant simultaneously (in 2 cases the variants 1+3a+3b, in 2 specimens the variants 1+3a and in 1 specimen the variant 1+3b). In one case of 96 EML4-ALK fusion gene and EGFR mutation were detected. All simultaneous genetic variants were confirmed using end-point PCR and direct sequencing. CONCLUSION: Our variant specific real time PCR assay is highly sensitive, fast, financially acceptable, applicable to FFPE and seems to be a valuable tool for the rapid prescreening of NSCLC patients in clinical practice, so, that most patients able to benefit from targeted therapy could be identified.
- MeSH
- adenokarcinom diagnóza genetika MeSH
- diagnostické techniky molekulární MeSH
- DNA sondy chemie MeSH
- fúzní onkogenní proteiny genetika metabolismus MeSH
- hybridizace in situ fluorescenční MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- messenger RNA genetika MeSH
- nádorové buněčné linie MeSH
- nádory plic diagnóza genetika MeSH
- nemalobuněčný karcinom plic diagnóza genetika MeSH
- spinocelulární karcinom diagnóza genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
