FLT3 Pathway Mutation PCR Array
|qBiomarker Somatic Mutation PCR Array: Human FLT3 Pathway
|Normal haematopoietic cells use complex systems to control proliferation,
differentiation, and cell death. Proliferation control is in part accomplished
through ligand-induced stimulation of receptor tyrosine kinases and their
downstream signaling cascades. Mutations in the FMS-like tyrosine receptor
kinase 3 (FLT3) gene have been found to be the most common genetic lesion in
acute myeloid leukaemia (AML), occurring in approximately 25% of cases. Recent
studies have established a link between FLT3 mutation status and response to
FLT3 inhibitors that warrants further investigation.
The Human FLT3 Pathway qBiomarker Somatic Mutation PCR Array is a
translational research tool that allows rapid and accurate profiling of the
somatic mutation status of the FLT3 gene and key downstream signaling
components. The utility of individual and multiple somatic mutation status
information in identifying key signaling transduction disruptions has been
demonstrated in numerous research studies. For example, the mutation status of
the EGFR and KRAS genes can predict the physiological response to certain drugs
targeting these molecules. The FLT3 Pathway qBiomarker Somatic Mutation PCR
Array, with its comprehensive content coverage, is designed for studying
mutations in the context of the FLT3 pathway and has the potential for
discovering and verifying drug target biomarkers for a variety of human cancers
involving the FLT3 signaling pathway and downstream effectors. This array covers
85 DNA sequence mutation assays designed to detect the most frequent,
functionally verified, and biologically most significant mutations in the FLT3
pathway. These mutations were chosen from curated, comprehensive somatic
mutation databases and peer-reviewed scientific literature. The simplicity of
the product format and operating procedure allows routine somatic mutation
profiling in any research laboratory with access to real-time PCR instruments.
The qBiomarker Somatic Mutation PCR Arrays are intended for molecular biology
applications. This product is not intended for the diagnosis, prevention, or
treatment of a disease.
| Modify this Array
19 mutation assays are included in this panel to cover the most
frequently identified FLT3 mutations. These include point mutations,
insertion and deletion mutations in the juxtamembrane region of the
coding sequence and in the activation loop portion of the sequence.
An extracellular domain mutation p.S451F and an N-terminal kinase
domain mutation A680V were also covered.
The panel includes an assay for the best known AKT1 mutation,
c.49G>A, p.E17K. This is a PH domain mutation that results in
constitutive targeting of AKT1 to plasma emebrane.
Two classes of mutation assays are included. One class covers
mutations that lead to increased BRAF kinase activity, such as the
p.L597 and p.V600 mutations. The other class detects mutations that
lead to impaired kinase activity, such as the p.G464V, p.G466V and
16 KRAS mutation assays provide comprehensive analysis capacity for
the most frequently occurring mutations in KRAS codon positions
12,13 and 61. Mutations at these positions result in reduced
intrinsic GTPase activity and/or cause KRAS unresponsive to RasGAP.
The p.L19F mutation assay is also included.
Similar to KRAS mutation assays, the >10 HRAS mutation assays on
this panel aim to cover the most important HRAS mutations identified
in cancers at codon 12, 13 and 61 positions.
12 NRAS mutation assays were included on the panel to cover codon
positions 12, 13 and 61, as well as mutation p.A18T.
4 assays for mutations with verified clinical significance in cancer
were included on the panel. These mutations cluster in MEK1
N-terminal negative regulatory domain and an adjacent domain, and
are all activating mutations (i.e. lead to upregulated intrinsic
MEK1 kinase activity).
PIK3CA (phosphatidylinositol 3-kinase catalytic subunit) gene:
The mutation assays covered on this panel can detect 7 most
frequently occurring PIK3CA mutations that belong to two classes:
p.H1047 mutations, which are activating, kinase domain mutations;
and mutations in P539-E545 region, which are helical domain
mutations that mimic activation by growth factors.
Included on the panel are 6 most commonly detected PTEN
loss-of-function mutations that are due to either truncation
(p.R233* and p.R130*) or point mutation-caused phosphatase
inactivation (p.R130 and p.R173 mutations).
Overview of the qBiomarker Somatic
Mutation PCR Array / Assay Protocol
Overview of the qBiomarker Somatic Mutation PCR Array / Assay
The procedure involves DNA extraction (QIAGEN QIAamp DNA Mini Kit or FFPE Tissue
Kit is recommended), an optional amplification (QIAGEN REPLI-g kit or REPLI-g
UltraFast kit is recommended) step for DNA isolated from fresh samples, qPCR
detection on qBiomarker Somatic Mutation PCR Arrays or Assays, and data analysis
(using the qBiomarker Somatic Mutation Data Analysis Template). An optional DNA
sample QC step immediately before the detection array or assay setup allows the
user to qualify the DNA samples.
Principle of Mutant Discrimination with ARMS®
Back to Top
- Prognostic significance of FLT3 internal tandem duplication and tyrosine
kinase domain mutations in acute promyelocytic leukemia: a systematic
review. Beitinjaneh A, Jang S, Roukoz H, Majhail NS. Leuk Res. 2010 Jul;
34(7):831-6. Epub 2010 Jan 21.
- The PKB/AKT pathway in cancer.Carnero A. Curr Pharm Des. 2010 Jan;
- BRAF, a target in melanoma: implications for solid tumor drug development.
Flaherty KT, McArthur G. Cancer. 2010 Jul 13. [Epub ahead of print].
- Clinical relevance of KRAS in human cancers. Jancík S, Drábek J,
Radzioch D, Hajdúch M.
J Biomed Biotechnol. 2010; 2010:150960.
- Oncogenic Ras in tumour progression and metastasis. Giehl K. Biol Chem.
2005 Mar; 386(3):193-205.
- MEK1 mutations confer resistance to MEK and B-RAF inhibition. Emery CM,
Vijayendran KG, Zipser MC, Sawyer AM, Niu L, Kim JJ, Hatton C, Chopra R,
Oberholzer PA, Karpova MB, MacConaill LE, Zhang J, Gray NS, Sellers WR,
Dummer R, Garraway LA. Proc Natl Acad Sci U S A. 2009 Dec 1;106(48):20411-6.
- PIK3CA mutations in human solid tumors: role in sensitivity to various
therapeutic approaches. Ligresti G, Militello L, Steelman LS, Cavallaro A,
Basile F, Nicoletti F, Stivala F, McCubrey JA, Libra M. Cell Cycle. 2009 May
- Oncogenic mutations as predictive factors in colorectal cancer. Ličvre A,
Blons H, Laurent-Puig P. Oncogene. 2010 May 27; 29(21):3033-43.
- PI(3)King Apart PTEN's Role in Cancer. Zhang S, Yu D. Clin Cancer Res.
2010 Jul 8. [Epub ahead of print].