Initial Intermittent Microwave Irradiation for Fluorescence In Situ Hybridization Analysis in Paraffin-Embedded Tissue Sections of Gastrointestinal Neoplasia

Kitayama, Yasuhiko; Igarashi, Hisaki; Sugimura, Haruhiko

doi:10.1038/labinvest.3780081

Cited by 37 publications

(38 citation statements)

References 6 publications

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“…Paraffin-embedded tissue sections were de-waxed and re-hydrated, and then boiled in 0.01 M citrate buffer (pH 6.0) to release closed chromosomal structures [15][16][17]. The sections were then hybridized with a Cy3-labeled BAC clone (RP11-339F13) for the EGFR locus (Advanced GenoTechs Co., Tsukuba, Japan) and an FITC-labeled probe for the centromere locus on chromosome 7, centromere enumeration probe (CEP) 7 (Vysis, Downer Groves, IL).…”

Section: Fluorescence In Situ Hybridization (Fish) Analysismentioning

confidence: 99%

Inverse relationship between the length of the EGFR CA repeat polymorphism in lung carcinoma and protein expression of EGFR in the carcinoma

Suzuki

Kageyama

Shinmura

et al. 2008

Journal of Surgical Oncology

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Section: Fluorescence In Situ Hybridization (Fish) Analysismentioning

confidence: 99%

Inverse relationship between the length of the EGFR CA repeat polymorphism in lung carcinoma and protein expression of EGFR in the carcinoma

Suzuki

Kageyama

Shinmura

et al. 2008

Journal of Surgical Oncology

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“…13 In brief, 4 mm sections were deparaffinized by five successive, 3-min washes in xylene followed by five washes in ethanol and heating in a 0.01 M citrate buffer (pH 6.0) using a microwave processor (MI-77, Azumaya Company, Tokyo, Japan) for 10 min. After treatment in 0.2% pepsin/0.01 N HCl for 10 min at 371C, the samples were aged in 0.1% NP-40/2 Â SSC for 10 min at 371C and their DNA was denatured by treatment in 70% formamide/2 Â SSC for 5 min at 851C.…”

Section: Fishmentioning

confidence: 99%

Protein overexpression and gene amplification of HER-2 and EGFR in colorectal cancers: an immunohistochemical and fluorescent in situ hybridization study

et al. 2004

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Overexpression of HER-2 and the epidermal growth factor receptor (EGFR) has been observed in many cancers, sometimes accompanied by gene amplification. To assess whether novel chemotherapies targeting these overexpressed proteins may be effective for the treatment of colorectal cancers, we examined the exact frequency of HER-2 and EGFR overexpression, the relationship between gene amplification and protein expression, and the heterogeneity of gene amplification within and between primary and metastatic tumors. We evaluated 244 colorectal cancers immunohistochemically. All tumors found to overexpress HER-2 or EGFR were further analyzed for gene amplification by fluorescent in situ DNA hybridization. Overexpression of HER-2 and EGFR was found in 8 (3%) and 19 (8%) of the 244 colorectal carcinomas, respectively. Gene amplification was observed in 100 and 58% of the tumors exhibiting HER-2 and EGFR overexpression, respectively. HER-2 amplification in cancer cells was characterized by clusters of hybridization signals, suggesting amplicons in homogeneously staining regions that were predominant in most primary and metastatic tumors. EGFR amplification, observed as scattered signals reminiscent of amplicons in double minute chromosomes, or coamplification of EGFR with the centromeric regions was observed as a minor population within primary tumors, and found in variety of populations in metastatic tumors. Overexpression of HER-2 and EGFR were observed in only a small fraction of colorectal carcinomas, but were frequently accompanied by gene amplification. The HER-2 gene is located on chromosomal region 17q11.2-q12 and the EGFR gene is located on 7p12. These encode 185 kDa (p185) and 170 kDa plasma membrane glycoproteins, respectively. They share approximately 50% overall homology and are composed of an N-terminus extracellular ligand-binding domain, a transmembrane lipophilic segment, and a C-terminus intracellular region containing a tyrosine kinase domain.

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“…This new protocol is especially versatile in application for formalin-fixed paraffin-embedded tissue specimens (Kitayama et al, 1999(Kitayama et al, , 2000bKobayashi et al, 2000;Lan et al, 1996;Sibony et al, 1995;Song et al, 2002). In this study, we applied this modified FISH technique using a panel of 18 chromosomal centromeric ␣-satellite probes and two region-specific probes to a chromosome-wide search for CNA in formalin-fixed paraffin-embedded tissue specimens from 51 cases of gastric cancer.…”

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confidence: 99%

Nonrandom Chromosomal Numerical Abnormality Predicting Prognosis of Gastric Cancer: A Retrospective Study of 51 Cases Using Pathology Archives

Kitayama

Igarashi

Watanabe

et al. 2003

Laboratory Investigation

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SUMMARY:Chromosomal or centromerical numerical abnormality (CNA) is a well-known characteristic of human cancer, but the extensive and specific documentation of CNA in gastric cancer is still sparse, partly because of difficulty in obtaining cytogenetic information. Taking advantage of a recently developed fluorescence in situ hybridization protocol for formalin-fixed paraffin-embedded tissues, we investigated CNA of 51 gastric cancer cases with a panel of 18 chromosome-specific ␣-satellite probes (for chromosomes 1-4, 6 -12, 15-18, 20, X and Y) and region specific probes (c-myc and p53) to enumerate respective chromosome numbers in interphase nuclei. The involved chromosomes exhibiting CNA were nonrandom in gastric cancer. Aberrations of chromosomes 1, 8, 17, 20, and X were frequent regardless of histologic types, whereas aberrations chromosomes 10, 15, and 18 occurred less often (p Ͻ 0.001). From a histopathologic standpoint, the mucocellular type had stable CNA in comparison with the tubular type (mucocellular type vs tubular type carcinoma: 21.0 Ϯ 10.63% vs 62.8 Ϯ 12.79%, p Ͻ 0.001). Interestingly, there was less extensive CNA in women (men vs women: 54.3 Ϯ 9.49% versus 24.9 Ϯ 12.23%, p Ͻ 0.001). A dramatic difference in the outcome was detected according to the involvement of chromosomes 3, 10, 11, 12, 17, and Y; that is, the cases with CNA of these chromosomes had worse prognosis. (Lab Invest 2003, 83:1311-1320.

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Initial Intermittent Microwave Irradiation for Fluorescence In Situ Hybridization Analysis in Paraffin-Embedded Tissue Sections of Gastrointestinal Neoplasia

Cited by 37 publications

References 6 publications

Inverse relationship between the length of the EGFR CA repeat polymorphism in lung carcinoma and protein expression of EGFR in the carcinoma

Inverse relationship between the length of the EGFR CA repeat polymorphism in lung carcinoma and protein expression of EGFR in the carcinoma

Protein overexpression and gene amplification of HER-2 and EGFR in colorectal cancers: an immunohistochemical and fluorescent in situ hybridization study

Nonrandom Chromosomal Numerical Abnormality Predicting Prognosis of Gastric Cancer: A Retrospective Study of 51 Cases Using Pathology Archives

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