Sequence mutations and amplification of PIK3CA and AKT2 genes in purified ovarian serous neoplasms

Kentaro Nakayama, Naomi Nakayama, Robert J Kurman, Leslie Cope, Gudrun Pohl, Yardena Samuels, Victor E Velculescu, Tian-Li Wang, Ie Ming Shih

Research output: Contribution to journalArticle

Abstract

Sequence mutations and gene amplifications lead to activation of the PIK3CA-AKT2 signaling pathway and have been reported in several types of neoplasms including ovarian cancer. Analysis of such genetic alterations, however, is usually complicated by contamination of normal cell DNA, artifacts associated with formalin-fixed tissues and the sensitivity of the techniques employed. In this study, we analyzed the sequence mutations in PIK3CA and AKT2 genes using purified tumor cells that were isolated from high-grade ovarian serous carcinomas and serous borderline tumors (SBTs) and assessed gene amplification using a dual-color FISH on tissue microarrays. Somatic sequence mutations in the kinase domain of AKT2 were not detected in any of the 65 ovarian tumors analyzed. Mutations of PIK3CA were rare, occurring only in one (2.3%) of 44 high-grade serous carcinomas and in only one (4.8%) of 21 SBTs. Dual-color FISH demonstrated that PIK3CA and AKT2 were not amplified in SBTs but amplified in 13.3% and 18.2% high-grade carcinomas, respectively. High-level amplification (>3 fold) was more frequently observed in AKT2 than in PIK3CA. Unlike mutations in ERBB2, KRAS and BRAF which are mutually exclusive in SBTs, coamplification of PIK3CA and AKT2 was present in five high-grade carcinomas including the OVCAR3 cells. Amplification in either of the genes occurred in 27% high-grade serous carcinomas. In conclusion, the methods we employed provide unambiguous evidence that somatic sequence mutations of PIK3CA and ATK2 are rare in ovarian serous tumors but amplification of both genes may play an important role in the development of high-grade ovarian serous carcinoma.

Original languageEnglish (US)
Pages (from-to)779-785
Number of pages7
JournalCancer Biology and Therapy
Volume5
Issue number7
StatePublished - Jul 2006

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Ovarian Neoplasms
Mutation
Carcinoma
Genes
Gene Amplification
Neoplasms
Color
Artifacts
Formaldehyde
Phosphotransferases
DNA

Keywords

  • Amplification
  • Digital karyotyping
  • FISH
  • Mutation
  • Ovarian cancer

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Sequence mutations and amplification of PIK3CA and AKT2 genes in purified ovarian serous neoplasms. / Nakayama, Kentaro; Nakayama, Naomi; Kurman, Robert J; Cope, Leslie; Pohl, Gudrun; Samuels, Yardena; Velculescu, Victor E; Wang, Tian-Li; Shih, Ie Ming.

In: Cancer Biology and Therapy, Vol. 5, No. 7, 07.2006, p. 779-785.

Research output: Contribution to journalArticle

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abstract = "Sequence mutations and gene amplifications lead to activation of the PIK3CA-AKT2 signaling pathway and have been reported in several types of neoplasms including ovarian cancer. Analysis of such genetic alterations, however, is usually complicated by contamination of normal cell DNA, artifacts associated with formalin-fixed tissues and the sensitivity of the techniques employed. In this study, we analyzed the sequence mutations in PIK3CA and AKT2 genes using purified tumor cells that were isolated from high-grade ovarian serous carcinomas and serous borderline tumors (SBTs) and assessed gene amplification using a dual-color FISH on tissue microarrays. Somatic sequence mutations in the kinase domain of AKT2 were not detected in any of the 65 ovarian tumors analyzed. Mutations of PIK3CA were rare, occurring only in one (2.3{\%}) of 44 high-grade serous carcinomas and in only one (4.8{\%}) of 21 SBTs. Dual-color FISH demonstrated that PIK3CA and AKT2 were not amplified in SBTs but amplified in 13.3{\%} and 18.2{\%} high-grade carcinomas, respectively. High-level amplification (>3 fold) was more frequently observed in AKT2 than in PIK3CA. Unlike mutations in ERBB2, KRAS and BRAF which are mutually exclusive in SBTs, coamplification of PIK3CA and AKT2 was present in five high-grade carcinomas including the OVCAR3 cells. Amplification in either of the genes occurred in 27{\%} high-grade serous carcinomas. In conclusion, the methods we employed provide unambiguous evidence that somatic sequence mutations of PIK3CA and ATK2 are rare in ovarian serous tumors but amplification of both genes may play an important role in the development of high-grade ovarian serous carcinoma.",
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