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PI3K-PTEN Pathway Mutation PCR Array

Human
 
qBiomarker Somatic Mutation PCR Array: Human PI3K-PTEN
The Human PI3K-PTEN Pathway qBiomarker Somatic Mutation PCR Array is a translational research tool that allows rapid and accurate profiling of the somatic mutation status for key genes in the PI3K-PTEN pathway: AKT1, AKT3, PIK3CA, PIK3R1, PIK3R5, PTEN, and STK11/LKB1. Components in this pathway are frequently mutated in human cancers (mutated in >30% solid tumor types) and therefore warrant extensive investigation to enhance the understanding of carcinogenesis and identify potential drug targets. 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 Human PI3K-PTEN Pathway qBiomarker Somatic Mutation PCR Array, with its comprehensive content coverage, is designed for studying mutations in the context of the PI3K-PTEN pathway and provides the potential to discover and verify drug target biomarkers for a variety of human cancers involving the PI3K-PTEN signaling pathway and downstream effectors. This array includes 87 DNA sequence mutation assays designed to detect the most frequent, functionally verified, and biologically significant mutations in the PI3K-PTEN 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.

 

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AKT genes:
The mutation assays detects the best known AKT mutation, c.49G>A, p.E17K (AKT1; a PH domain mutation that results in constitutive targeting of AKT1 to plasma membrane), the activating mutations c.145G>A, p.E49K (AKT1) and c.511G>A, p.G171R (AKT3).

PIK3CA (phosphatidylinositol 3-kinase catalytic subunit) gene:
The mutation assays covered on this panel can detect 37 most frequently occurring PIK3CA mutations that are mainly clustered in the kinase domain (activating mutations) and in the P539-E545 region and vicinity, which are helical domain mutations that mimic activation by growth factors. The assays allow a comprehensive profiling of PIK3CA mutations that have been identified in a variety of cancers.

PIK3R1 and PIK3R5 (PI3K regulatory subunit) genes:
Two recurrent disinhibitory (c.1126G>A, p.G376R and c.1690A>G, p.N564D) mutations in PIK3R1 and a recurrent oncogenic mutation in PIK3R5 are included.

PTEN gene:
Included on the panel are 33 most commonly detected PTEN loss-of-function mutations that are mainly due to truncation or point mutation-caused phosphatase inactivation.

STK11/LKB1 gene:
Included on the panel are 11 most commonly detected STK11/LKB1 inactivation mutations that are mainly due to truncation or point mutation.

 

Assay Functional Annotations How It Works References Resources
 

Overview of the qBiomarker Somatic Mutation PCR Array / Assay Protocol

Overview of the qBiomarker Somatic Mutation PCR Array / Assay Protocol.
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®

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Assay Functional Annotations How It Works References Resources
 
  1. The PKB/AKT pathway in cancer.Carnero A. Curr Pharm Des. 2010 Jan; 16(1):34-44
  2. BRAF, a target in melanoma: implications for solid tumor drug development. Flaherty KT, McArthur G. Cancer. 2010 Jul 13. [Epub ahead of print]
  3. Clinical relevance of KRAS in human cancers. Jancík S, Drábek J, Radzioch D, Hajdúch M. J Biomed Biotechnol. 2010; 2010:150960.
  4. Oncogenic Ras in tumour progression and metastasis. Giehl K. Biol Chem. 2005 Mar; 386(3):193-205
  5. 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
  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 1; 8(9):1352-8
  7. Oncogenic mutations as predictive factors in colorectal cancer. Ličvre A, Blons H, Laurent-Puig P. Oncogene. 2010 May 27; 29(21):3033-43
  8.  PI(3)King Apart PTEN's Role in Cancer. Zhang S, Yu D. Clin Cancer Res. 2010 Jul 8. [Epub ahead of print]
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Assay Functional Annotations How It Works References Resources
 

User Manual qBiomarker Somatic Mutation PCR Array System (PDF)
Data Analysis qBiomarker Somatic Mutation PCR Array Data Analysis Software
Application Data Detection Limits, Cancer Pathways
FAQ Frequently Asked Questions about Somatic Mutation Assays and Arrays
Webinar qBiomarker Somatic Mutation Analysis: Real-World Application Data
Slide Presentation> Presentation about qBiomarker Somatic Mutation Assays and Arrays (PDF)
Scientific Poster A Novel Tool for Pathway-Focused Cancer Mutation Profiling (PDF)
Presented at American Association for Cancer Research 2011
White Paper Rapid and accurate cancer somatic mutation profiling with the qBiomarker Somatic Mutation PCR Array (PDF)
Product Profile For screening biology-focused panels of gene mutations (PDF)

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