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Comparative Study
. 2014 Aug 9:9:153.
doi: 10.1186/s13000-014-0153-4.

Diagnostic application of PIK3CA mutation analysis in Chinese esophageal cancer patients

Zizhen Ming, Dongxian Jiang, Qin Hu, Xiaojing Li, Jie Huang, Yifan Xu, Yalan Liu, Chen Xu, Xiuguo Hua  1 , Yingyong Hou
Affiliations
Comparative Study

Diagnostic application of PIK3CA mutation analysis in Chinese esophageal cancer patients

Zizhen Ming et al. Diagn Pathol. .

Abstract

Background: The PIK3CA gene mutation was found to associate with prognosis and might affect molecular targeted therapy in esophageal carcinoma (EC). The aim of this study is to compare different methods for analyzing the PIK3CA gene mutation in EC.

Methods: Genomic DNA was extracted from 106 surgically resected EC patient tissues. The PIK3CA mutation status (exons 9 and 20) were screened by mutant-enrich liquid chip (ME-Liquidchip), Sanger sequencing, and pyrosequencing. And all samples with mutations were independently reassessed using amplification refractory mutation system (ARMS) methods again.

Results: PIK3CA mutation rates were identified as 11.3% (12/106) by ME-Liquidchip. 10 mutations occurred in exon 9 and 2 in exon 20, including G1624A:E542K (n = 4), G1633A:E545K (n = 6) and A3140G:H1047R (n = 2). The results were further verified by ARMS methods. Among these 12 cases characterized for PIK3CA mutation, however, only 7 and 6 cases were identified by Sanger sequencing (6.6%,7/106) and pyrosequencing (5.7%,6/106), respectively.

Conclusion: Sanger sequencing and pyrosequencing are less sensitive and are not efficiently applicable to the detection of PIK3CA mutation in EC samples. Choosing between ME-Liquidchip and ARMS will depend on laboratory facilities and expertise.

Virtual slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_153.

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Figures

Figure 1
Figure 1
Representative results for PIK3CA exon 9 and 20 mutation analysis. EC058 (A) and EC037 (B) were detected harboring E545K and H1047R mutation using Sanger sequencing and ARMS, but the same samples were considered wt according to pyrosequencing.
See this image and copyright information in PMC

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