Proliferating cell nuclear antigen (PCNA): a new marker to study human colonic cell proliferation.

Kubben, F. J. G. M.; Peeters-Haesevoets, A.; Engels, L.G.J.B.; Baeten, C.G.; Schütte, B.; Arends, Jan Willem; Stockbrügger, R.W.; Blijham, Geert H.

doi:10.1136/gut.35.4.530

Cited by 169 publications

(118 citation statements)

References 27 publications

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“…For the chromoendoscopy study, 48 ACF were analyzed by real-time PCR (see Supplementary Table S2). PCNA is induced by EGFR and widely used as a marker of cell proliferation (34,35). Because expression levels of PCNA mRNA and protein levels correlate closely (36), we used PCNA mRNA abundance to assess the proliferative status of ACF analyzed at the RNA level.…”

Section: Resultsmentioning

confidence: 99%

Epidermal Growth Factor Receptor Signaling Is Up-regulated in Human Colonic Aberrant Crypt Foci

et al. 2006

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Aberrant crypt foci (ACF) are collections of abnormal colonic crypts with heterogeneous molecular and pathologic characteristics. Large and dysplastic ACF are putative precursors of colon cancer with neoplastic risk related to increased proliferation. In this study, we examined the role of epidermal growth factor receptor (EGFR) signaling in regulating ACF proliferation. Using magnification chromoendoscopy, we collected large ACF with endoscopic features of dysplasia and separately biopsied adjacent mucosa. Transcript levels were measured by real-time PCR, proteins were assessed by Western blotting, and levels were expressed as fold changes of adjacent mucosa. K-ras and B-Raf mutations were assessed by PCR and Ras activation by the ratio Ras-GTP / (Ras-GTP + Ras-GDP). At the RNA level, 38% of ACF were hyperproliferative, with proliferating cell nuclear antigen (PCNA) mRNA z2-fold of adjacent mucosa. Hyperproliferative ACF had significantly increased mRNA levels of EGFR (6.0 F 1.7-fold), transforming growth factor-A (14.4 F 5.0-fold), heparin-binding EGF-like growth factor (4.5 F 1.4-fold), cyclin D1 (4.6 F 0.7-fold), and cyclooxygenase-2 (COX-2; 9.3 F 4.2-fold; P < 0.05). At the protein level, 46% of ACF were hyperproliferative (PCNA, 3.2 F 1.2-fold). In hyperproliferative ACF, 44% possessed significant increases in four EGFR signaling components: EGFR (9.5 F 1.3-fold), phosphoactive ErbB2 (2.6 F 0.4-fold), phosphoactive extracellular signal-regulated kinase (3.7 F 1.1-fold), and cyclin D1 (3.4 F 0.8-fold; P < 0.05). Ras was activated in 46% of ACF (3.2 F 0.4-fold; P < 0.05), but K-ras mutations were present in only 7% of ACF. In contrast to COX-2 mRNA, the protein was not increased in hyperproliferative ACF. In summary, we have shown that ACF with up-regulated PCNA possess increased EGFR signaling components that likely contribute to the enhanced proliferative state of

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Section: Resultsmentioning

confidence: 99%

Epidermal Growth Factor Receptor Signaling Is Up-regulated in Human Colonic Aberrant Crypt Foci

et al. 2006

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“…PCNA and Ki67 are abundant proteins in the colon with restricted detection at the bottom of the crypts. Correlated with their localization, Ki67 and PCNA play a role in proliferation (15,20,32): Ki67, although universally used as a proliferative marker, still has an unknown function; PCNA is a member of the DNA sliding clamp family and plays a role not only in proliferation through DNA replication but also in DNA repair (26). The fact that both Ki67 and PCNA were enhanced 2 days after microbiota transfer illustrated a rapid and strong boost in proliferation.…”

Section: Discussionmentioning

confidence: 99%

Microbiota matures colonic epithelium through a coordinated induction of cell cycle-related proteins in gnotobiotic rat

Cherbuy

Honvo-Houeto

Bruneau

et al. 2010

American Journal of Physiology-Gastrointestinal and Liver Physi

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Cherbuy C, Honvo-Houeto E, Bruneau A, Bridonneau C, Mayeur C, Duée PH, Langella P, Thomas M. Microbiota matures colonic epithelium through a coordinated induction of cell cycle-related proteins in gnotobiotic rat. Am J Physiol Gastrointest Liver Physiol 299: G348 -G357, 2010. First published May 13, 2010; doi:10.1152/ajpgi.00384.2009.-Previous studies have suggested that intestinal microbiota modulates colonic epithelium renewal. The objective of our work was to study the effects of microbiota on colonic epithelium structure and cell cycle-related proteins by using gnotobiotic rats. Colonic crypts and amount of cell cycle-related proteins were compared between germ-free (GF), conventional (CV), and conventionalized rats by histochemistry and Western blot. Ki67 and proliferating cell nuclear antigen (PCNA) were used as surrogates for proliferative cells; p21 cip1 and p27 kip1 were markers of cell cycle arrest; anti-and proapoptotic proteins, Bcl2 and Bax, respectively, were also studied. We observed 40% increase of the crypt proliferative area 2 days after inoculation of GF rats with a complex microbiota. This recruitment of proliferative cells may account for the 30% increase of crypt depth observed between CV and GF rats. The hyperproliferative boost induced by microbiota was compensated by a fourfold increase of p21 cip1 and p27 kip1 involved in cell cycle arrest and a 30% drop of antiapoptotic Bcl2 protein while Bax was unchanged. Inductions of p21 cip1 , p27 kip1 , and PCNA protein were not paralleled by an increase of the corresponding mRNA. We also showed that p21 cip1 induction by microbiota was partially restored by Bacteroides thetaiotaomicron, Ruminococcus gnavus, and Clostridium paraputrificum. Colonization of the colon by a complex microbiota increases the crypt depth of colon epithelium. This event takes place in conjunction with a multistep process: a hyperproliferative boost accompanied by compensatory events as induction of p21 cip1 and p27 kip1 and decrease of Bcl2.intestine; germ-free; PCNA; Bcl2; p21 cip1 ; p27 kip1 THE GASTROINTESTINAL TRACT of mammals is colonized by a community of microorganisms reaching levels as high as 10 11 bacteria/g of content in the colon. The intestinal microbiota includes more than 1,000 different species of bacteria belonging mainly to two divisions, the Firmicutes and the Bacteroidetes. Microbiota represents a large genomic repertoire as revealed by metagenomic approaches (12) and an efficacious organ to recover energy (33). Intestinal microbiota has coevolved with its host, exploiting mutualistic interactions (24). Thus the gut immune tolerance toward microbiota partly involves fine-tuned regulation of the transcription factor NF-B by commensal bacteria (19). Furthermore, microbiota induce expression of fucosylated cell surface glycans by mammalian gut epithelium (4, 29), which may be used as receptors for bacterial adhesins or as energy sources for the intestinal bacteria, especially in limited nutrient availability (45). Finally, our group has shown that in...

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“…Colons were harvested, fixed, and paraffin embedded according to standard techniques. Sections of colon tissue were dewaxed and stained for PCNA with a monoclonal anti-mouse primary antibody (Dakopatts, Copenhagen, Denmark) and a goat anti-mouse IgG-horseradish peroxidase secondary antibody (Dakopatts) with diaminobenzidine as substrate as described previously (15). The labeling index was calculated as the number of immunopositive cells multiplied by 100 and divided by the total number of cells per colonic crypt.…”

Section: Methodsmentioning

confidence: 99%

Stimulation of proliferation in the colorectal mucosa by gastrin precursors is blocked by desferrioxamine

Ferrand

Lachal

Bramante

et al. 2010

American Journal of Physiology-Gastrointestinal and Liver Physi

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Precursors of the peptide hormone gastrin stimulate proliferation in the colorectal mucosa and promote the development of colorectal carcinoma. Gastrins bind two ferric ions selectively and with high affinity, and the biological activity of glycine-extended gastrin (Ggly) in vitro is dependent on the presence of ferric ions. The aim of the present study was to determine whether or not iron is required for biological activity of progastrin and Ggly in vivo. Rats that had undergone a colostomy were infused with Ggly, and proliferation was measured in the defunctioned rectal mucosa. Proliferation was also measured in the colonic mucosa of hGAS and MTI-Ggly mice, which, by definition, overexpress progastrin and Ggly, respectively. The requirement for iron was assessed by thrice-weekly injection of the chelating agent desferrioxamine (DFO). The proliferation index in the defunctioned rectal mucosa was significantly increased in the Ggly-infused rats, and the increase was significantly reduced after treatment with DFO. Treatment with DFO significantly reduced the crypt height and proliferation index in the colonic mucosa of hGAS and MTI-Ggly mice but had no effect on the same variables in wild-type mice. These observations are consistent with the hypothesis that the biological activity of progastrin and Ggly in vivo is dependent on the presence of ferric ions and further suggest that chelating agents may block the stimulatory effects of gastrin precursors in the development of colorectal carcinoma.

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Proliferating cell nuclear antigen (PCNA): a new marker to study human colonic cell proliferation.

Cited by 169 publications

References 27 publications

Epidermal Growth Factor Receptor Signaling Is Up-regulated in Human Colonic Aberrant Crypt Foci

Epidermal Growth Factor Receptor Signaling Is Up-regulated in Human Colonic Aberrant Crypt Foci

Microbiota matures colonic epithelium through a coordinated induction of cell cycle-related proteins in gnotobiotic rat

Stimulation of proliferation in the colorectal mucosa by gastrin precursors is blocked by desferrioxamine

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