A novel ex vivo method for assaying adhesion of cancer cells to the peritoneum

Asao, Takayuki; Yazawa, Shin; Kudo, Shigeharu; Takenoshita, S; Nagamachi, Yukio

doi:10.1016/0304-3835(94)90031-0

Cited by 22 publications

(19 citation statements)

References 13 publications

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“…For the inhibition experiments, the cells were pre-treated with neutralizing antibody to different integrin subunits at 37°C for 30 min prior to their being incubated with the section of peritoneum. This procedures was described before by Asao et al (25).…”

Section: Methodsmentioning

confidence: 99%

Elevated Expression of Cyr61 Enhances Peritoneal Dissemination of Gastric Cancer Cells through Integrin α2β1

Lin∥

Chang

Lin

et al. 2007

Journal of Biological Chemistry

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Cysteine-rich 61 (Cyr61/CCN1) is involved in human gastric cancer development and progression. Nonetheless, the role of Cyr61 as regards peritoneal dissemination of such cancers has not yet been completely characterized. We used liposome-mediated transfection to establish Cyr61, or antisense Cyr61, expression vectors into gastric cancer AGS or MKN45 cell lines. Transfectants were tested by means of a cancer-cell adhesion assay in vitro and ex vivo. Furthermore, a functional integrin fluorescence-activated cell sorting assay, reverse transcription-PCR, and an AP-1 reporter assay were performed to investigate the potential signaling pathway of Cyr61. It was shown that stable transfection of Cyr61 into the AGS cell line strongly enhanced its adhesion ability. The overexpression of Cyr61 within AGS cells significantly increased the functional expression of integrin ␣ 2 ␤ 1 . Function-neutralizing antibody to integrin ␣ 2 ␤ 1 effectively suppressed the Cyr61-mediated enhanced adhesion of AGS cells to peritoneal tissue. Promoter assays of integrin ␣2 gene further revealed that the AP-1 pathway was evidently activated within Cyr61-expressing AGS cells. Animal studies have revealed that mice injected with Cyr61-overexpressed AGS cells featured a greater number of peritoneal seeding nodules and a lower survival rate than the Neo control cell lines, and when such cells were treated with functional blocking antibody to integrin ␣ 2 ␤ 1 , they were able to elicit a decline in the peritoneal dissemination. The data suggest that Cyr61 may contribute to the peritoneal dissemination of gastric cancer by promoting tumor-cell adhesion ability through the up-regulation of the functional integrin ␣ 2 ␤ 1 via an AP-1-dependent pathway.

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Section: Methodsmentioning

confidence: 99%

Elevated Expression of Cyr61 Enhances Peritoneal Dissemination of Gastric Cancer Cells through Integrin α2β1

Lin∥

Chang

Lin

et al. 2007

Journal of Biological Chemistry

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“…Human gastric cancer cell lines MKN74 and MKN45, as well as human colon cancer cell lines COLO201, WiDr and COLO205 cells were provided from Japanese Cancer Research Resources Bank Corp (Tokyo, Japan). MKN45 and MKN74 cells were from poorly dierentiated and well-dierentiated types of gastric cancers, respectively (Asao et al, 1994). The blots were developed by a standard enhanced chemiluminescence (ECL) method (Amersham).…”

Section: Increased Apoptosis Resistance Of Mkn74 Cells Expressing Bclmentioning

confidence: 99%

“…For these experiments, bcl-2 and bag-1 cDNAs encoding either full-length Bcl-2 (Tsujimoto and Croce, 1986) or BAG-1 (Takayama et al, 1995) proteins were stably introduced into MKN74 cells. These gastric cancer cells are capable of dissemination in the murine peritoneal cavity (Asao et al, 1994). After selection in neomycin, the Bcl-2 or BAG-1 transfectant cells were analysed by immunoblotting using Bcl-2-and BAG-1-speci®c antibodies.…”

Section: Increased Apoptosis Resistance Of Mkn74 Cells Expressing Bclmentioning

confidence: 99%

Prolonged cell survival enhances peritoneal dissemination of gastric cancer cells

et al. 1998

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“…MKN45 and MKN74 cells were derived from poorly di erentiated and well-di erentiated types of gastric cancers, respectively (Asao et al, 1994). They were maintained in D-MEM supplemented with 10% FBS in a humidi®ed atmosphere of 5% CO 2 -air at 378C.…”

Section: Reagents and Cell Culturementioning

confidence: 99%