Gene expression analysis of distinct populations of cells isolated from mouse and human inner ear FFPE tissue using laser capture microdissection – a Technical report based on preliminary findings

Pagedar, Nitin A.; Wang, Wen; Chen, Daniel H.-C.; Davis, Roger T.; López, Iván; Wright, Charles G.; Alagramam, Kumar N.

doi:10.1016/j.brainres.2006.01.057

Cited by 28 publications

(23 citation statements)

References 26 publications

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“…Successful RNA extraction from microdissected FFPE samples has been shown to be feasible (47) and capable of producing results similar to frozen fixed tissue, although RNA quantity and quality are reduced (48). Five patients were selected with archival FFPE blocks from each of the three stages (CA, HGD, and QC) cut to 7 μm and mounted on glass slides.…”

Section: Methodsmentioning

confidence: 99%

Truncated neurokinin-1 receptor is increased in colonic epithelial cells from patients with colitis-associated cancer

Gillespie¹,

Leeman²,

Watts³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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Patients with chronic ulcerative colitis (UC) are at high risk for developing colorectal cancer. In this study, archival formalin-fixed paraffin-embedded colonic tissue from patients with UC who developed carcinoma (CA) or high-grade dysplasia (HGD) was examined for changes in expression of the proinflammatory and mitogenic neurokinin-1 receptor (NK-1R). Laser capture microscopy was used to microdissect epithelia from areas of colons that showed histologic evidence of CA, HGD, and epithelia that were not dysplastic or cancerous but did contain evidence of prior inflammation (quiescent colitis). mRNA was extracted from the dissected tissue, and PCR array analysis was performed on extracted mRNA. Two antibodies were necessary to separately estimate the protein levels of the truncated (tr-NK-1R) and full-length (fl-NK-1R) receptors by immunohistochemistry. mRNA expression of tr-NK-1R increased 14-fold (P = 0.02) when comparing the HGD and CA groups. In contrast, the fl-NK-1R transcript showed no significant differences among groups. The protein levels of the total NK-1R increased by 40% (P = 0.02) in HGD and 80% (P = 0.0007) in CA compared with quiescent colitis. There were no significant changes in protein levels of the fl-NK-1R. We conclude that the increase in total NK-1R protein in HGD and CA is attributable to an increase in tr-NK-1R, suggesting there may be a functional role for tr-NK-1R in malignant transformation in colitis-associated cancer. The tr-NK-1R could prove useful as a diagnostic marker to identify patients at risk for neoplasia and may serve as a useful therapeutic target in the treatment of colitisassociated cancer.colon cancer | receptor splice variants | substance P | IBD C hronic inflammation is associated with a higher rate of cancer development in various organs (1). More specifically, patients with ulcerative colitis (UC) are at high risk for developing colorectal cancer (2); both extent and duration of intestinal inflammation further increase risk (3, 4). These associations suggest that chronic intestinal inflammation is a causative factor in carcinogenesis in patients with UC, and that there is a causal link between chronic inflammation and increased risk of cancer. These observations raise the question of the signaling pathways by which long-standing inflammation might underlie the development of cancer.Substance P (SP) is a proinflammatory neuropeptide described by von Euler and Gaddum (5) and later isolated and characterized by Chang and Leeman (6). SP is synthesized by a variety of cells, most notably neurons and inflammatory cells such as macrophages (7). SP's primary receptor, the neurokinin-1 receptor (NK-1R), can mediate a variety of physiologic and pathophysiologic responses, including cell proliferation, migration, and inflammation (8). The NK-1R has been identified in epithelial cells in rodent models of colonic inflammation (9, 10) and in patients with UC as well as Crohn disease (11)(12)(13)(14). However, not all of these studies are in agreement regarding epithelial NK-1R e...

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Section: Methodsmentioning

confidence: 99%

Truncated neurokinin-1 receptor is increased in colonic epithelial cells from patients with colitis-associated cancer

Gillespie¹,

Leeman²,

Watts³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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show abstract

“…However, in the end meaningful microarray studies will need to be done on individual cells or homogeneous cell populations. The isolation of single cells is now possible using laser capture microdissection and this technique has been used to isolate cochlear and vestibular cells (Cristobal et al, 2004(Cristobal et al, , 2005Kimura et al, 2005;Pagedar et al, 2006).…”

Section: Inner Ear Cdna Librariesmentioning

confidence: 99%

Gene expression profiling analysis of the inner ear

et al. 2007

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“…Unstained frozen sections were used instead of fixed, paraffin embedded, stained sections for the LMPC, as the RNA stability of material gained from the former is considerably higher (Goldsworthy et al, 1999). Paraffin embedded and stained sections are also adequate for LMPC since identification of the target areas is actually much easier due to the tissue staining, but recovering usable RNA from them is a tedious task as demonstrated by Pagedar et al (2006). The two-tailed Mann-Whitney-test was used for statistical assessment of differential gene expression: *** p 6 0.001; n.s., not significant.…”

Section: Discussionmentioning

confidence: 99%

Technical report: Laser microdissection and pressure catapulting is superior to conventional manual dissection for isolating pure spiral ganglion fractions from the cochlea

et al. 2008

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Gene expression analysis of distinct populations of cells isolated from mouse and human inner ear FFPE tissue using laser capture microdissection – a Technical report based on preliminary findings

Cited by 28 publications

References 26 publications

Truncated neurokinin-1 receptor is increased in colonic epithelial cells from patients with colitis-associated cancer

Truncated neurokinin-1 receptor is increased in colonic epithelial cells from patients with colitis-associated cancer

Gene expression profiling analysis of the inner ear

Technical report: Laser microdissection and pressure catapulting is superior to conventional manual dissection for isolating pure spiral ganglion fractions from the cochlea

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