• Logo
  • SBMUJournals

High Resolution Melting Analysis for Rapid Detection of PIK3CA Gene Mutations in Bladder Cancer: A Mutated Target for Cancer Therapy

Zahra Ousati Ashtiani, Abdol Rasoul Mehrsai, Mohammad Reza Pourmand, Gholam Reza Pourmand




Purpose: PIK3CA gene mutations have clinical importance and their presence is associated with therapy response. They are also considered as a molecule for targeted therapy. As regards to their importance, genetic variation within a population as well as among different populations, this study was conducted to detect common mutations of exons 9 and 20 and other probable mutations in PIK3CA gene as well as their frequencies in Iranian bladder cancer patients.
Materials and methods: Paired tumor and adjacent normal tissues samples were obtained from 50 bladder cancer patients. Mutations of PIK3CA gene were detected using High Resolution Melting (HRM) analysis which is a
highly sensitive, repeatable, rapid, and cost-effective technique. To determine the precision of the HRM analysis, Sanger sequencing analysis was used.
Results: The result showed that mutations were present in 10% (5/50) of the subjects. The majority of these cases (4/5) had the mutation(s) in exon 9, spanning over five different mutations, among which three of them were actually novel mutations. Further analysis showed that 2 cases had simultaneous mutations for exon 9. In addition to novel mutations, the PIK3CA mutation rate observed in Iranian bladder patients was not as frequent as previous reports and COSMIC.
Conclusion: HRM can be used as a rapid and sensitive method for mutation screening. Dysregulation of PIK3CA gene in bladder cancer reveals its potentials as a mechanistic link for cancer development, which in turn suggests its special use in interventional studies for targeted therapy.


Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011; 61:69-90.

Salehi A, Khezri A, Malekmakan L, Aminsharifi A. Epidemiologic status of

bladder cancer in Shiraz, southern Iran. Asian Pac J Cancer Prev. 2011; 12:1323-7.

Rafiemanesh H, Rajaei-Behbahani N, Khani Y, Hosseini S. Incidence trend and epidemiology of common cancers in the center of Iran. Glob

J Health Sci. 2016; 8:146.

Castillo-Martin M, Domingo-Domenech J, Karni-Schmidt O, Matos T, Cordon-Cardo C Molecular pathways of urothelial development

and bladder tumorigenesis. Urol Oncol. 2010; 28:401-408.

Duenas M, Martínez‐Fernández M, García‐Escudero R, et al. PIK3CA gene alterations in bladder cancer are frequent and associate with reduced recurrence in non‐muscle invasive tumors. Mol Carcinog. 2015; 54:566-76.

Meeks J J, Herr H W. Office-based management of nonmuscle invasive bladder cancer. Urol Clin North Am. 2013; 40:473-9.

Goebell P J, Knowles M A. Bladder cancer or bladder cancers? Genetically distinct malignant conditions of the urothelium. Urol Oncol. 2010; 28: 409-428.

Yuan T, Cantley L. PI3K pathway alterations in cancer: variations on a theme. Oncogene 2008; 27:5497-510.

Platt FM, Hurst CD, Taylor CF, Gregory WM, Harnden P, Knowles MA. Spectrum of phosphatidylinositol 3-kinase pathway gene alterations in bladder cancer. Clin Can Res. 2009; 15:6008-17.

Knowles MA, Platt FM, Ross RL, Hurst CD. Phosphatidylinositol 3-kinase (PI3K) pathway activation in bladder cancer. Can Metas Rev. 2009; 28:305-16.

Kommineni N, Jamil K, Pingali U, Addala L, Naidu M. Association of PIK3CA gene mutations with head and neck squamous cell carcinomas. Neoplasma. 2014; 62: 72-80.

Kandula M, Chennaboina KK, Ammi R, Raju S. Phosphatidylinositol 3-kinase (PI3KCA) oncogene mutation analysis and gene expression profiling in primary breast cancer patients. Asian Pac J Cancer Prev. 2013;


Dirican E, Akkiprik M, Özer A. Mutation distributions and clinical correlations of PIK3CA gene mutations in breast cancer. Tumor Biol. 2016; 37: 7033-7045.

Guedes J G, Veiga I, Rocha P, et al. High resolution melting analysis of KRAS, BRAF and PIK3CA in KRAS exon 2 wild-type

metastatic colorectal cancer. BMC cancer. 2013; 13: 169.

Konopka B, Janiec–Jankowska A, Kwiatkowska E, et al. PIK3CA mutations and amplification in endometrioid endometrial carcinomas: relation to other genetic defects and clinicopathologic status of the tumors.

Hum Pathol. 2011; 42: 1710-19.

Tserga A, Chatziandreou I, Michalopoulos NV, Patsouris E, Saetta AA. Mutation of genes of the PI3K/AKT pathway in breast cancer supports their potential importance as biomarker for breast cancer aggressiveness.

Virchows Arch. 2016; 469: 35-43.

López-Knowles E, Hernández S, Malats N, et al. PIK3CA mutations are an early genetic alteration associated with FGFR3 mutations in superficial papillary bladder tumors. Cancer Res. 2006; 66: 7401-4.

Serizawa RR, Ralfkiær U, Steven K, et al. Integrated genetic and epigenetic analysis of bladder cancer reveals an additive diagnostic value of FGFR3 mutations and hypermethylation events. Int J Cancer. 2011;

: 78-87.

Kompier LC, Lurkin I, van der Aa MN, van Rhijn BW, van der Kwast TH, Zwarthoff EC. FGFR3, HRAS, KRAS, NRAS and PIK3CA mutations in bladder cancer and their potential as biomarkers for surveillance and therapy.

PloS one. 2010; 5: e13821.

Juanpere N, Agell L, Lorenzo M, et al. Mutations in FGFR3 and PIK3CA, singly or combined with RAS and AKT1, are associated with AKT but not with MAPK pathway activation in urothelial bladder cancer. Hum

Pathol. 2012; 43: 1573-82.

Agell L, Hernández S, Salido M, et al. PI3K signaling pathway is activated by PIK3CA mRNA overexpression and copy gain in prostate tumors, but PIK3CA, BRAF, KRAS and AKT1 mutations are infrequent events. Mod Path. 2011; 24: 443-52.

Vorkas PA, Poumpouridou N, Agelaki S, Kroupis C, Georgoulias V, Lianidou ES. PIK3CA hotspot mutation scanning by a novel and highly sensitive high resolution small amplicon melting analysis method. J Mol

Diagn. 2010; 12: 697-704.

Montgomery JL, Sanford LN, Wittwer CT. High-resolution DNA melting analysis in clinical research and diagnostics. Expert Rev Mol Diagn. 2010; 10: 219-40.

Nosho K, Kawasaki T, Longtine JA, et al. PIK3CA mutation in colorectal cancer: relationship with genetic and epigenetic alterations. Neoplasia. 2008; 10: 534-41.

Xing JC, Tufano RP, Murugan AK, et al. Single nucleotide polymorphism rs17849071 G/T in the PIK3CA gene is inversely associated

with follicular thyroid cancer and PIK3CA amplification. PloS one. 2012; 7: e49192.

Wong K K, Engelman J A, Cantley L C. Targeting the PI3K signaling pathway in cancer. Currt Opin Genet Dev. 2010; 20: 87-90.

Nisa L, Häfliger P, Poliaková M, et al. PIK3CA hotspot mutations differentially impact responses to MET targeting in METdriven and non-driven preclinical cancer models. Mol Cancer. 2017; 16: 93-106.

Millis SZ, Bryant D, Basu G, et al. Molecular profiling of infiltrating urothelial carcinoma of bladder and nonbladder origin. Clin Genitourin

Cancer 2015; 13:e37-e49

Azizi Tabesh G, Izadi P, Fereidooni F, Emami Razavi AN, Tavakkoly Bazzaz J. The High Frequency of PIK3CA Mutations in Iranian

Breast Cancer Patients. Cancer Inves. 2017; 35:36-42

Bonetti L R, Barresi V, Bettelli S, Caprera C, ManfredinI S, Maiorana A. Analysis of KRAS, NRAS, PIK3CA and BRAF mutational profile in poorly differentiated clusters (PDC) of KRAS mutated colon cancer. Hum Pathol.

; 62: 91-98

DOI: http://dx.doi.org/10.22037/uj.v0i0.3987


  • There are currently no refbacks.