Fluorescent labelling of intestinal epithelial cells reveals independent long-lived intestinal stem cells in a crypt

Horita, Nobukatsu; Tsuchiya, Kiichiro; Hayashi, Ryohei; Fukushima, Kunihiro; Hibiya, Shuji; Fukuda, Masayoshi; Kano, Yoshihito; Mizutani, Tomohiro; Nemoto, Yasuhiro; Yui, Satoru; Okamoto, Ryuichi; Nakamura, Toshio; Watanabe, Mamoru

doi:10.1016/j.bbrc.2014.10.091

Cited by 10 publications

(6 citation statements)

References 22 publications

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“…Sporadic human colon cancer‐derived DLD1 cells were cultured as described previously . Plasmid DNA was transfected by lentivirus infection, as described previously . Blasticidin (7.5 μg/mL, Invitrogen, Carlsbad, CA, USA) was used only in the selection of transfected cells.…”

Section: Methodsmentioning

confidence: 99%

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Atonal homolog 1 protein stabilized by tumor necrosis factor α induces high malignant potential in colon cancer cell line

et al. 2015

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Patients with inflammatory bowel disease (IBD) have an increased risk of developing colitis-associated colorectal cancer (CAC). CAC cells often develop chemoresistance, resulting in a poorer prognosis than that of sporadic colorectal cancer (CRC). The mechanism by which CAC enhances malignant potential remains unknown. We have previously reported that the proteasomal degradation of the transcription factor Atonal homolog 1 (Atoh1) protein results in the non-mucinous form of CRC. It also remains unknown whether Atoh1 protein is expressed in CAC. Therefore, in the present study, we investigated whether Atoh1 protein stabilizes in CAC. Consequently, the treatment with TNF-α stabilized Atoh1 protein through the inactivation of GSK-3β via Akt, resulting in the mucinous form of CRC cell lines. Atoh1 protein also enriched cancer stem cells with upregulated Lgr5 expression and cells in G0/G1 cell cycle phase, resulting in both the chemoresistance to 5-fluorouracil and oxaliplatin and the promotion of cell migration. Immunofluorescence of the human mucinous CAC specimens showed the accumulation of NF-κB p65 at nuclei with the expression of Atoh1 in mucinous cancer. In conclusion, the inflammation associated with carcinogenesis may preserve the differentiation system of intestinal epithelial cell (IEC), resulting in the acquisition of both the mucinous phenotype and high malignant potential associated with the enrichment of cancer stem cell.

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Section: Methodsmentioning

confidence: 99%

“…(13) Plasmid DNA was transfected by lentivirus infection, as described previously. (14) Blasticidin (7.5 lg ⁄ mL, Invitrogen, Carlsbad, CA, USA) was used only in the selection of transfected cells. Blasticidin was excluded during the treatment with reagents.…”

Section: Methodsmentioning

confidence: 99%

Atonal homolog 1 protein stabilized by tumor necrosis factor α induces high malignant potential in colon cancer cell line

et al. 2015

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“…A major advantage of this system is the ability to greatly expand both tissue-specific stem cells and their differentiated progeny from very limited amounts of starting material such as biopsies, facilitating in-depth analyses of stem cell behaviour, drug screening, disease modelling and genetic screening. Indeed, the intestinal organoids have already been used extensively for analysing stem cell behaviour, identifying niche components, modelling pathogen-epithelia interactions, gene editing, disease modelling and orthotopic transplantation 2,4,[17][18][19][20][21][53][54][55][56][57][58][59][60][61][62][63][64] (Fig. 2).…”

Section: Applications Of Organoid Technologymentioning

confidence: 99%

Organoids as an in vitro model of human development and disease

2016

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The in vitro organoid model is a major technological breakthrough that has already been established as an essential tool in many basic biology and clinical applications. This near-physiological 3D model facilitates an accurate study of a range of in vivo biological processes including tissue renewal, stem cell/niche functions and tissue responses to drugs, mutation or damage. In this Review, we discuss the current achievements, challenges and potential applications of this technique.

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“…Primary antibodies were visualized using secondary antibodies conjugated to either Alexa-594 or Alexa-488 (Life Technologies). Sections were mounted using VectaShield mounting medium containing DAPI (Vector Laboratories) and visualized by confocal laser fluorescent microscopy (FLUOVIEW FV10i;Olympus) as previously described (19).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Reduced Human α-defensin 6 in Noninflamed Jejunal Tissue of Patients with Crohn’s Disease

Hayashi

Tsuchiya

Fukushima

et al. 2016

Inflammatory Bowel Diseases

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Fluorescent labelling of intestinal epithelial cells reveals independent long-lived intestinal stem cells in a crypt

Abstract: The use of our original lentiviral transgene system to mark individual organoid crypt stem cells showed that long-lived plural crypt stem cells might independently serve as intestinal epithelial cells, resulting in the formation of a completely functional villus.

Cited by 10 publications

References 22 publications

Atonal homolog 1 protein stabilized by tumor necrosis factor α induces high malignant potential in colon cancer cell line

Atonal homolog 1 protein stabilized by tumor necrosis factor α induces high malignant potential in colon cancer cell line

Organoids as an in vitro model of human development and disease

Reduced Human α-defensin 6 in Noninflamed Jejunal Tissue of Patients with Crohn’s Disease

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