Analysis of proliferation, apoptosis and keratin expression in cultured normal and immortalized human buccal keratinocytes

Hansson, Annette; Bloor, Balvinder K.; Sarang, Zsolt; Haig, Ylva; Morgan, Peter; Stark, Hans Jürgen; Fusenig, Norbert E.; Ekstrand, Jan; Grafström, Roland C.

doi:10.1034/j.1600-0722.2003.00010.x

Cited by 22 publications

(30 citation statements)

References 38 publications

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“…This was in line with previous experiments in melanoma cells (21). A spontaneous induction of Ck's 8 and 18 expression could be shown in SV40T-immortalized buccal mucosa cells, in tobacco induced leukoplakia and after introduction of v-H-Ras in epidermal mouse keratinocytes (22). The involvement of Ck 8/18 in SCC progression could further be demonstrated in chemically induced SCCs in the mouse skin.…”

Section: Discussionmentioning

confidence: 68%

Cytokeratin alteration in oral leukoplakia and oral squamous cell carcinoma

Fillies

Jogschies²,

Kleinheinz³

et al. 2007

Oncol Rep

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Section: Discussionmentioning

confidence: 68%

Cytokeratin alteration in oral leukoplakia and oral squamous cell carcinoma

Fillies

Jogschies²,

Kleinheinz³

et al. 2007

Oncol Rep

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“…In oral carcinomas, the proportion of active caspase‐3‐positive cells with apoptotic morphology was markedly higher than in normal oral epithelium, confirming the findings of Macluskey et al 26 that the apoptotic index is increased during oral carcinogenesis. Hansson et al 27 showed that in organotypic culture, the immortalized buccal epithelial cell line, SVpgC2a, consistently exhibited a greater apoptotic index than normal oral keratinocytes. We found that active caspase‐3 staining facilitated the scoring of apoptotic index in oral tissue.…”

Section: Discussionmentioning

confidence: 99%

Caspase‐3 expression is reduced, in the absence of cleavage, in terminally differentiated normal oral epithelium but is increased in oral squamous cell carcinomas and correlates with tumour stage

Hague

Eveson

MacFarlane

et al. 2004

The Journal of Pathology

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Oral carcinomas are known to have a greater apoptotic index than normal oral epithelium, evident as shrinking cells with condensed chromatin. In this study, these morphologically apoptotic cells stained positively for cleaved (active) caspase-3. In normal oral epithelium, cleaved caspase-3 positive-cells were only rarely detected. The terminally differentiated surface epithelial layers did not express cleaved caspase-3. The caspase-3 pro-enzyme showed a gradient of expression in normal oral epithelium, decreasing with differentiation. No expression was detectable in surface epithelial layers. Lack of expression of the major 'executioner' caspase-3 may, at least in part, explain differences in morphology between terminally differentiated and apoptotic cells. In cancers of different tissue origins, caspase-3 pro-enzyme expression can be either increased or decreased compared with normal tissue counterparts. To determine how caspase-3 expression alters during oral carcinogenesis, caspase-3 expression was compared in 39 samples of normal oral epithelium and 54 oral squamous cell carcinomas. Squamous cell carcinomas had more intense caspase-3 staining than normal epithelium (p < 0.001). Moreover, within the oral squamous cell carcinoma series, there was significantly more intense nuclear and cytoplasmic staining with increasing STNMP stage (p = 0.017 and 0.03, respectively). This was a reflection of significant associations with site (S), palpable lymph nodes (N), and differentiation (P). Both caspase-3 staining intensity and the percentage of cells positive for caspase-3 were inversely associated with differentiation. Studies of the mechanisms by which high levels of caspase-3 expression are tolerated in oral carcinoma cells may identify targets that can be used to harness caspase-3 overexpression for therapeutic benefit.

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“…Keratinocytes were cultured on dermal equivalents composed of type I collagen and fibroblasts and raised to the air-medium interface to promote differentiation. This model has also been applied to keratinocytes derived from the oral cavity, including buccal (16,17,33,34,73), lingual (38), and peritonsillar (62) surfaces. Several of these studies underscored the importance of fibroblasts located in adjacent connective tissue, which provide permissive and instructive signals to differentiating keratinocytes (64,77).…”

Section: Discussionmentioning

confidence: 99%

Activation of Kaposi's Sarcoma-Associated Herpesvirus Lytic Gene Expression during Epithelial Differentiation

Johnson¹,

Maronian

Vieira³

2005

J Virol

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The oral cavity has been identified as the major site for the shedding of infectious Kaposi's sarcomaassociated herpesvirus (KSHV). While KSHV DNA is frequently detected in the saliva of KSHV seropositive persons, it does not appear to replicate in salivary glands. Some viruses employ the process of epithelial differentiation for productive viral replication. To test if KSHV utilizes the differentiation of oral epithelium as a mechanism for the activation of lytic replication and virus production, we developed an organotypic raft culture model of epithelium using keratinocytes from human tonsils. This system produced a nonkeratinized stratified squamous oral epithelium in vitro, as demonstrated by the presence of nucleated cells at the apical surface; the expression of involucrin and keratins 6, 13, 14, and 19; and the absence of keratin 1. The activation of KSHV lytic-gene expression was examined in this system using rKSHV.219, a recombinant virus that expresses the green fluorescent protein during latency from the cellular EF-1␣ promoter and the red fluorescent protein (RFP) during lytic replication from the viral early PAN promoter. Infection of keratinocytes with rKSHV.219 resulted in latent infection; however, when these keratinocytes differentiated into a multilayered epithelium, lytic cycle activation of rKSHV.219 occurred, as evidenced by RFP expression, the expression of the late virion protein open reading frame K8.1, and the production of infectious rKSHV.219 at the epithelial surface. These findings demonstrate that KSHV lytic activation occurs as keratinocytes differentiate into a mature epithelium, and it may be responsible for the presence of infectious KSHV in saliva.

show abstract

Analysis of proliferation, apoptosis and keratin expression in cultured normal and immortalized human buccal keratinocytes

Cited by 22 publications

References 38 publications

Cytokeratin alteration in oral leukoplakia and oral squamous cell carcinoma

Cytokeratin alteration in oral leukoplakia and oral squamous cell carcinoma

Caspase‐3 expression is reduced, in the absence of cleavage, in terminally differentiated normal oral epithelium but is increased in oral squamous cell carcinomas and correlates with tumour stage

Activation of Kaposi's Sarcoma-Associated Herpesvirus Lytic Gene Expression during Epithelial Differentiation

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