Fluorescence in situ hybridization analysis with a tissue microarray: ‘FISH and chips’ analysis of pathology archives

Sugimura, Haruhiko; Mori, Hiroki; Nagura, Kiyoko; Kiyose, Shinichiro; Hong, Tao; Isozaki, Masaru; Igarashi, Hisaki; Shinmura, Kazuya; Hasegawa, Akio; Kitayama, Yasuhiko; Tanioka, Fumihiko

doi:10.1111/j.1440-1827.2010.02561.x

Cited by 28 publications

(28 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…50 lg of cell lysates were add to the reaction buffer (80 mM Tissue microarray (TMA), immunohistochemistry and western blotting. Details on TMA construction and immunohistological evaluation are based on previous literature (23) and in the Supporting Information. Briefly, the expression of LPCAT4 in CRC was categorized according to the modified algorithm in previous literature, that is, the staining intensities were categorized as follows: blue (0); blue-brown (1); brown (2); and bright brown (3).…”

Section: Methodsmentioning

confidence: 99%

Accumulated phosphatidylcholine (16:0/16:1) in human colorectal cancer; possible involvement of LPCAT4

et al. 2013

Self Cite

View full text Add to dashboard Cite

The identification of cancer biomarkers is critical for target-linked cancer therapy. The overall level of phosphatidylcholine (PC) is elevated in colorectal cancer (CRC). To investigate which species of PC is overexpressed in colorectal cancer, an imaging mass spectrometry was performed using a panel of non-neoplastic mucosal and CRC tissues. In the present study, we identified a novel biomarker, PC(16:0 ⁄ 16:1), in CRC using imaging mass spectrometry. Specifically, elevated levels of PC(16:0 ⁄ 16:1) expression were observed in the more advanced stage of CRC. Our data further showed that PC(16:0 ⁄ 16:1) was specifically localized in the cancer region when examined using imaging mass spectrometry. Notably, because the ratio of PC(16:0 ⁄ 16:1) to lyso-PC(16:0) was higher in CRC, we postulated that lyso-PC acyltransferase (LPCAT) activity is elevated in CRC. In an in vitro analysis, we showed that LPCAT4 is involved in the deregulation of PC (16: (1) Previous studies have reported that CRC contains increased amounts of phospholipids as well as an altered phospholipid composition of the CRC cell membrane.(2,3) These changes in membrane phospholipid levels can affect cell proliferation, viability and tumor development.(2,4) Moreover, although phosphatidylcholine (PC) is the most dominant phospholipid in both non-neoplastic and cancer tissues,the amount of PC is highly increased in CRC cells.(2) In addition, the changes in membrane potential and the increased PC ⁄ phosphatidylethanolamine (PE) composition rate are related to the grade of CRC malignancy.(3) In higher eukaryotes, PC is synthesized via two pathways: (i) the triple methylation of PE; and (ii) the cytidine diphosphate (CDP)-choline pathway. (6) However, the steady-state composition of PC species is maintained by the remodeling cycle (Lands' cycle).(7) The precise and concerted deacylation by phospholipase A 2 (PLA 2 ) and reacylation by lyso-PC acyltransferase (LPCAT) are required for normal cell functioning. Notably, the elevated expression of LPCAT1 was associated with colon cancer growth. (8) LPCAT1 is a member of the LPCAT family (LPCAT1-4) and shows LPCAT activity, preferentially incorporating palmitate into PC.(8) However, the relationship between PC remodeling and the progression of cancer, as well as the class of LPCAT involved in this process, remains unclear.Direct mass spectrometry (MS) of biological tissue sections using matrix-assisted laser desorption ⁄ ionization (MALDI) can profile many molecules including phospholipid subtypes. (9) Furthermore, this approach can be extended to imaging MS, which can visualize the distribution of biomolecules in the tissue section.(10-13) Because specific antibodies against lipids and macromolecules are often difficult to obtain, MALDI imaging is a suitable option for detecting distinct species of these molecules directly in a tissue section. This technique has already been applied to various human cancers including prostate and gastric cancer. (14,15) Although this emerging analytical technique was initial...

show abstract

Section: Methodsmentioning

confidence: 99%

Accumulated phosphatidylcholine (16:0/16:1) in human colorectal cancer; possible involvement of LPCAT4

et al. 2013

Self Cite

View full text Add to dashboard Cite

show abstract

“…Clinicopathological information on EPHB3 alone (not accompanied with EPHB2) in human gastrointestinal tract cancers remains limited. A clinical interpretation of the over-expression and/or amplification of EPHB3 (Figure 1) in gastrointestinal cancer [70,71] awaits further investigations.…”

Section: Ephb3mentioning

confidence: 99%

EPH-EPHRIN in human gastrointestinal cancers

Sugimura

Wang²,

Mori³

et al. 2010

WJGO

Self Cite

View full text Add to dashboard Cite

Ever since its discovery two decades ago, the erythropoietin-producing hepatoma (EPH)-EPHRIN system has been shown to play multifaceted roles in human gastroenterological cancer as well as neurodevelopment. Overexpression, amplification and point mutations have been found in many human cancers and many investigators have shown correlations between these up-regulations and tumor angiogenesis. Thus, the genes in this family are considered to be potential targets of cancer therapy. On the other hand, the down-regulation of some members as a result of epigenetic changes has also been reported in some cancers. Furthermore, the correlation between altered expressions and clinical prognosis seems to be inconclusive. A huge amount of protein-protein interaction studies on the EPH-EPHRIN system have provided a basic scheme for signal transductions, especially bi-directional signaling involving EPH-ERPHRIN molecules at the cell membrane. This information also provides a manipulative strategy for harnessing the actions of these molecules. In this review, we summarize the known alterations of EPH-EPHRIN genes in human tumors of the esophagus, stomach, colorectum, liver and pancreas and present the perspective that the EPH-EPHRIN system could be a potential target of cancer therapy.

show abstract

“…Human archival tissue blocks contain specimens of human tumors in various stages of development, which are precious in the post-human-genome-sequencing era. Based on their findings and other's work, Sugimura et al (Sugimura, Mori et al 2010) stated that the intensive application BAC clones as probes for FISH that have exact 'addresses' in the whole genome will become a useful diagnostic tools for pathologists. Thousands of BAC clones are commercially available, and any of them can be used as FISH probes.…”

Section: Fluorescence In Situ Hybridization (Fish) Analysis Of Patholmentioning

confidence: 99%

“…Thousands of BAC clones are commercially available, and any of them can be used as FISH probes. Sugimura et al tested 100 BAC probes containing different kinase loci in a gastric, colorectal, and lung cancer detection sets (20 cases for each organ) by using tissue microarray (TMA)-FISH technology (Sugimura, Mori et al 2010). Sugimura et al found that unexpected kinase loci were amplified in a significant proportion of human common solid tumors (Sugimura, Mori et al 2010).…”

Section: Fluorescence In Situ Hybridization (Fish) Analysis Of Patholmentioning

confidence: 99%

Gene Functional Studies Using Bacterial Artificial Chromosome (BACs)

Liu¹,

Guo²,

Battle³

et al. 2011

Bacterial Artificial Chromosomes

View full text Add to dashboard Cite

Fluorescence in situ hybridization analysis with a tissue microarray: ‘FISH and chips’ analysis of pathology archives

Cited by 28 publications

References 64 publications

Accumulated phosphatidylcholine (16:0/16:1) in human colorectal cancer; possible involvement of LPCAT4

Accumulated phosphatidylcholine (16:0/16:1) in human colorectal cancer; possible involvement of LPCAT4

EPH-EPHRIN in human gastrointestinal cancers

Gene Functional Studies Using Bacterial Artificial Chromosome (BACs)

Contact Info

Product

Resources

About