Limits...
Enhancive effects of Lewis y antigen on CD44-mediated adhesion and spreading of human ovarian cancer cell line RMG-I.

Gao L, Yan L, Lin B, Gao J, Liang X, Wang Y, Liu J, Zhang S, Iwamori M - J. Exp. Clin. Cancer Res. (2011)

Bottom Line: Western Blot confirmed that the content of CD44 in RMG-I-H cells was 1.46 times of that in RMG-I cells.The co-location of Lewis y antigen and CD44 was confirmed by co-immunoprecipitation.The mRNA level of CD44 in both cell lines was similar (P > 0.05).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital Affiliated to China Medical University, Shenyang, 110004, P R of China.

ABSTRACT

Background: This study aimed to investigate the molecular structural relationship between cell adhesive molecule CD44 and Lewis y antigen, and determine the effects of Lewis y antigen on CD44-mediated adhesion and spreading of ovarian cancer cell line RMG-I and the Lewis y antigen-overexpressed cell line RMG-I-H.

Methods: The expression of CD44 in RMG-I and RMG-I-H cells before and after treatment of Lewis y monoclonal antibody was detected by immunocytochemistry; the expression of Lewis y antigen and CD44 was detected by Western Blot. The structural relationship between Lewis y antigen and CD44 was determined by immunoprecipitation and confocal laser scanning microscopy. The adhesion and spreading of RMG-I and RMG-I-H cells on hyaluronic acid (HA) were observed. The expression of CD44 mRNA in RMG-I and RMG-I-H cells was detected by real-time RT-PCR.

Results: Immunocytochemistry revealed that the expression of CD44 was significantly higher in RMG-I-H cells than in RMG-I cells (P < 0.01), and its expression in both cell lines was significantly decreased after treatment of Lewis y monoclonal antibody (both P < 0.01). Western Blot confirmed that the content of CD44 in RMG-I-H cells was 1.46 times of that in RMG-I cells. The co-location of Lewis y antigen and CD44 was confirmed by co-immunoprecipitation. The co-expression of CD44 and Lewis y antigen in RMG-I-H cells was 2.24 times of that in RMG-I cells. The adhesion and spreading of RMG-I-H cells on HA were significantly enhanced as compared to those of RMG-I cells (P < 0.01), and this enhancement was inhibited by Lewis y monoclonal antibody (P < 0.01). The mRNA level of CD44 in both cell lines was similar (P > 0.05).

Conclusion: Lewis y antigen strengthens CD44-mediated adhesion and spreading of ovarian cancer cells.

Show MeSH

Related in: MedlinePlus

Co-location of CD44 and Lewis y antigen on RMG-I-H cells observed under confocal laser scanning microscope. Red fluoscence on the upper left panel indicates CD44 expression; green fluoscence on the upper right panel indicates Lewis y antigen expression; blue fluoscence on the upper right panel indicates cell nuclear location; the lower right panel is a merged image of the other three panels. Lewis y antigen CD44 mainly expressed in the cell membrane observed under the confocal laser scanning microscope, and it were seen as yellow fluorescence after the two overlap, suggesting that Lewis y antigen and CD44 co-localizated in the cell membrane.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3045975&req=5

Figure 2: Co-location of CD44 and Lewis y antigen on RMG-I-H cells observed under confocal laser scanning microscope. Red fluoscence on the upper left panel indicates CD44 expression; green fluoscence on the upper right panel indicates Lewis y antigen expression; blue fluoscence on the upper right panel indicates cell nuclear location; the lower right panel is a merged image of the other three panels. Lewis y antigen CD44 mainly expressed in the cell membrane observed under the confocal laser scanning microscope, and it were seen as yellow fluorescence after the two overlap, suggesting that Lewis y antigen and CD44 co-localizated in the cell membrane.

Mentions: Under the confocal laser scanning microscope, CD44 presented red fluoscence mainly on cell membrane and partly in cytoplasm; Lewis y antigen presented green fluoscence mainly on cell membrane (Figure 2). Both red fluoscence and green fluoscence were accumulated at the margin of cell clusters and overlapped as yellow fluoscence, indicating the co-location of CD44 and Lewis y antigen.


Enhancive effects of Lewis y antigen on CD44-mediated adhesion and spreading of human ovarian cancer cell line RMG-I.

Gao L, Yan L, Lin B, Gao J, Liang X, Wang Y, Liu J, Zhang S, Iwamori M - J. Exp. Clin. Cancer Res. (2011)

Co-location of CD44 and Lewis y antigen on RMG-I-H cells observed under confocal laser scanning microscope. Red fluoscence on the upper left panel indicates CD44 expression; green fluoscence on the upper right panel indicates Lewis y antigen expression; blue fluoscence on the upper right panel indicates cell nuclear location; the lower right panel is a merged image of the other three panels. Lewis y antigen CD44 mainly expressed in the cell membrane observed under the confocal laser scanning microscope, and it were seen as yellow fluorescence after the two overlap, suggesting that Lewis y antigen and CD44 co-localizated in the cell membrane.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3045975&req=5

Figure 2: Co-location of CD44 and Lewis y antigen on RMG-I-H cells observed under confocal laser scanning microscope. Red fluoscence on the upper left panel indicates CD44 expression; green fluoscence on the upper right panel indicates Lewis y antigen expression; blue fluoscence on the upper right panel indicates cell nuclear location; the lower right panel is a merged image of the other three panels. Lewis y antigen CD44 mainly expressed in the cell membrane observed under the confocal laser scanning microscope, and it were seen as yellow fluorescence after the two overlap, suggesting that Lewis y antigen and CD44 co-localizated in the cell membrane.
Mentions: Under the confocal laser scanning microscope, CD44 presented red fluoscence mainly on cell membrane and partly in cytoplasm; Lewis y antigen presented green fluoscence mainly on cell membrane (Figure 2). Both red fluoscence and green fluoscence were accumulated at the margin of cell clusters and overlapped as yellow fluoscence, indicating the co-location of CD44 and Lewis y antigen.

Bottom Line: Western Blot confirmed that the content of CD44 in RMG-I-H cells was 1.46 times of that in RMG-I cells.The co-location of Lewis y antigen and CD44 was confirmed by co-immunoprecipitation.The mRNA level of CD44 in both cell lines was similar (P > 0.05).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital Affiliated to China Medical University, Shenyang, 110004, P R of China.

ABSTRACT

Background: This study aimed to investigate the molecular structural relationship between cell adhesive molecule CD44 and Lewis y antigen, and determine the effects of Lewis y antigen on CD44-mediated adhesion and spreading of ovarian cancer cell line RMG-I and the Lewis y antigen-overexpressed cell line RMG-I-H.

Methods: The expression of CD44 in RMG-I and RMG-I-H cells before and after treatment of Lewis y monoclonal antibody was detected by immunocytochemistry; the expression of Lewis y antigen and CD44 was detected by Western Blot. The structural relationship between Lewis y antigen and CD44 was determined by immunoprecipitation and confocal laser scanning microscopy. The adhesion and spreading of RMG-I and RMG-I-H cells on hyaluronic acid (HA) were observed. The expression of CD44 mRNA in RMG-I and RMG-I-H cells was detected by real-time RT-PCR.

Results: Immunocytochemistry revealed that the expression of CD44 was significantly higher in RMG-I-H cells than in RMG-I cells (P < 0.01), and its expression in both cell lines was significantly decreased after treatment of Lewis y monoclonal antibody (both P < 0.01). Western Blot confirmed that the content of CD44 in RMG-I-H cells was 1.46 times of that in RMG-I cells. The co-location of Lewis y antigen and CD44 was confirmed by co-immunoprecipitation. The co-expression of CD44 and Lewis y antigen in RMG-I-H cells was 2.24 times of that in RMG-I cells. The adhesion and spreading of RMG-I-H cells on HA were significantly enhanced as compared to those of RMG-I cells (P < 0.01), and this enhancement was inhibited by Lewis y monoclonal antibody (P < 0.01). The mRNA level of CD44 in both cell lines was similar (P > 0.05).

Conclusion: Lewis y antigen strengthens CD44-mediated adhesion and spreading of ovarian cancer cells.

Show MeSH
Related in: MedlinePlus