Limits...
A Novel Function of CD82/KAI1 in Sialyl Lewis Antigen-Mediated Adhesion of Cancer Cells: Evidence for an Anti-Metastasis Effect by Down-Regulation of Sialyl Lewis Antigens.

Yoshihama N, Yamaguchi K, Chigita S, Mine M, Abe M, Ishii K, Kobayashi Y, Akimoto N, Mori Y, Sugiura T - PLoS ONE (2015)

Bottom Line: In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells.Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses.We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

View Article: PubMed Central - PubMed

Affiliation: Section of Oral and Maxillofacial Surgery, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.

ABSTRACT
We have recently elucidated a novel function for CD82 in E-cadherin-mediated homocellular adhesion; due to this function, it can inhibit cancer cell dissociation from the primary cancer nest and limit metastasis. However, the effect of CD82 on selectin ligand-mediated heterocellular adhesion has not yet been elucidated. In this study, we focused on the effects of the metastasis suppressor CD82/KAI1 on heterocellular adhesion of cancer cells to the endothelium of blood vessels in order to further elucidate the function of tetraspanins. The over-expression of CD82 in cancer cells led to the inhibition of experimentally induced lung metastases in mice and significantly inhibited the adhesion of these cells to human umbilical vein epithelial cells (HUVECs) in vitro. Pre-treatment of the cells with function-perturbing antibodies against sLea/x significantly inhibited the adhesion of CD82-negative cells to HUVECs. In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells. Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses. Knockdown of ST3GAL4 on CD82-negative wild-type cells inhibited expression of sLex and reduced cell adhesion to HUVECs. We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

No MeSH data available.


Related in: MedlinePlus

Inhibitory effect of CD82 on cancer cell adhesion to a HUVEC monolayer culture.Fluorescently labeled h1299/zeo or h1299/CD82 cells (4.0 × 104 cells/well) were applied to a HUVEC monolayer culture and allowed to adhere at 37°C. Adhered cells were quantified after 30 min. Adhered h1299/zeo (A) and h1299/CD82 (B) cells and the effect of function-perturbing antibodies on cell adhesion to the HUVEC monolayer were analysed (C). h1299 cells were pre-treated with 5 μg/ml of the indicated antibodies and then applied to the HUVEC monolayer. Experiments were performed in triplicate and the number of adhered cells was averaged. Bars indicate the standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4414583&req=5

pone.0124743.g002: Inhibitory effect of CD82 on cancer cell adhesion to a HUVEC monolayer culture.Fluorescently labeled h1299/zeo or h1299/CD82 cells (4.0 × 104 cells/well) were applied to a HUVEC monolayer culture and allowed to adhere at 37°C. Adhered cells were quantified after 30 min. Adhered h1299/zeo (A) and h1299/CD82 (B) cells and the effect of function-perturbing antibodies on cell adhesion to the HUVEC monolayer were analysed (C). h1299 cells were pre-treated with 5 μg/ml of the indicated antibodies and then applied to the HUVEC monolayer. Experiments were performed in triplicate and the number of adhered cells was averaged. Bars indicate the standard deviation.

Mentions: The adhesion of cancer cells to blood vessels involves the interaction of P- and E-selectin on the epithelial cells of blood vessels with the corresponding sLex and sLea carbohydrate ligands on the surface of cancer cells [12]. We analysed the effect of CD82 on cell adhesion to blood vessels using a previously described cell adhesion assay in HUVECs [11]. It was previously confirmed that HUVECs express both P- and E-selectin [11]. We observed that approximately 65% of the loaded h1299/zeo cells adhered to the HUVEC monolayer. Conversely, h1299/CD82 cells showed significantly less cell adhesion, with only 6.6% adherence to the HUVEC monolayer (Fig 2A and 2B). Pre-treatment with function-perturbing antibodies against sLex, sLea, or β1 integrins inhibited h1299/zeo adhesion to HUVECs by 9.4%, 8.4%, and 30.2%, respectively (Fig 2C). In contrast, function-perturbing antibodies against sLex/a showed no significant inhibition of h1299/CD82 cell adhesion to HUVECs. These results strongly suggested an inhibitory role for CD82 in sialyl Lewis-mediated cellular adhesion.


A Novel Function of CD82/KAI1 in Sialyl Lewis Antigen-Mediated Adhesion of Cancer Cells: Evidence for an Anti-Metastasis Effect by Down-Regulation of Sialyl Lewis Antigens.

Yoshihama N, Yamaguchi K, Chigita S, Mine M, Abe M, Ishii K, Kobayashi Y, Akimoto N, Mori Y, Sugiura T - PLoS ONE (2015)

Inhibitory effect of CD82 on cancer cell adhesion to a HUVEC monolayer culture.Fluorescently labeled h1299/zeo or h1299/CD82 cells (4.0 × 104 cells/well) were applied to a HUVEC monolayer culture and allowed to adhere at 37°C. Adhered cells were quantified after 30 min. Adhered h1299/zeo (A) and h1299/CD82 (B) cells and the effect of function-perturbing antibodies on cell adhesion to the HUVEC monolayer were analysed (C). h1299 cells were pre-treated with 5 μg/ml of the indicated antibodies and then applied to the HUVEC monolayer. Experiments were performed in triplicate and the number of adhered cells was averaged. Bars indicate the standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414583&req=5

pone.0124743.g002: Inhibitory effect of CD82 on cancer cell adhesion to a HUVEC monolayer culture.Fluorescently labeled h1299/zeo or h1299/CD82 cells (4.0 × 104 cells/well) were applied to a HUVEC monolayer culture and allowed to adhere at 37°C. Adhered cells were quantified after 30 min. Adhered h1299/zeo (A) and h1299/CD82 (B) cells and the effect of function-perturbing antibodies on cell adhesion to the HUVEC monolayer were analysed (C). h1299 cells were pre-treated with 5 μg/ml of the indicated antibodies and then applied to the HUVEC monolayer. Experiments were performed in triplicate and the number of adhered cells was averaged. Bars indicate the standard deviation.
Mentions: The adhesion of cancer cells to blood vessels involves the interaction of P- and E-selectin on the epithelial cells of blood vessels with the corresponding sLex and sLea carbohydrate ligands on the surface of cancer cells [12]. We analysed the effect of CD82 on cell adhesion to blood vessels using a previously described cell adhesion assay in HUVECs [11]. It was previously confirmed that HUVECs express both P- and E-selectin [11]. We observed that approximately 65% of the loaded h1299/zeo cells adhered to the HUVEC monolayer. Conversely, h1299/CD82 cells showed significantly less cell adhesion, with only 6.6% adherence to the HUVEC monolayer (Fig 2A and 2B). Pre-treatment with function-perturbing antibodies against sLex, sLea, or β1 integrins inhibited h1299/zeo adhesion to HUVECs by 9.4%, 8.4%, and 30.2%, respectively (Fig 2C). In contrast, function-perturbing antibodies against sLex/a showed no significant inhibition of h1299/CD82 cell adhesion to HUVECs. These results strongly suggested an inhibitory role for CD82 in sialyl Lewis-mediated cellular adhesion.

Bottom Line: In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells.Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses.We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

View Article: PubMed Central - PubMed

Affiliation: Section of Oral and Maxillofacial Surgery, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.

ABSTRACT
We have recently elucidated a novel function for CD82 in E-cadherin-mediated homocellular adhesion; due to this function, it can inhibit cancer cell dissociation from the primary cancer nest and limit metastasis. However, the effect of CD82 on selectin ligand-mediated heterocellular adhesion has not yet been elucidated. In this study, we focused on the effects of the metastasis suppressor CD82/KAI1 on heterocellular adhesion of cancer cells to the endothelium of blood vessels in order to further elucidate the function of tetraspanins. The over-expression of CD82 in cancer cells led to the inhibition of experimentally induced lung metastases in mice and significantly inhibited the adhesion of these cells to human umbilical vein epithelial cells (HUVECs) in vitro. Pre-treatment of the cells with function-perturbing antibodies against sLea/x significantly inhibited the adhesion of CD82-negative cells to HUVECs. In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells. Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses. Knockdown of ST3GAL4 on CD82-negative wild-type cells inhibited expression of sLex and reduced cell adhesion to HUVECs. We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

No MeSH data available.


Related in: MedlinePlus