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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).

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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.

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The mRNA expression of CD44 and α1, 2-FT in RMG-I and RMG-I-H cells were tested by quantitative Real-Time RT-PCR. The mRNA level of α1, 2-FT was significantly increased, but the mRNA level of CD44 was almost the same in RMG-1-hFUT cells and RMG-1 cells. (**P < 0.01, * P > 0.05).
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Figure 4: The mRNA expression of CD44 and α1, 2-FT in RMG-I and RMG-I-H cells were tested by quantitative Real-Time RT-PCR. The mRNA level of α1, 2-FT was significantly increased, but the mRNA level of CD44 was almost the same in RMG-1-hFUT cells and RMG-1 cells. (**P < 0.01, * P > 0.05).

Mentions: The 2-ΔΔCT value of mRNA level of CD44 in RMG-I-H cells is 79% of that in RMG-I cells, which had no significant difference (P > 0.05), whereas the mRNA level of α1,2-FT in RMG-I-H cells was increased by 3.07 times of that in RMG-I cells detected by Real-time PCR (P < 0.01). (Figure 4).


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)

The mRNA expression of CD44 and α1, 2-FT in RMG-I and RMG-I-H cells were tested by quantitative Real-Time RT-PCR. The mRNA level of α1, 2-FT was significantly increased, but the mRNA level of CD44 was almost the same in RMG-1-hFUT cells and RMG-1 cells. (**P < 0.01, * P > 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: The mRNA expression of CD44 and α1, 2-FT in RMG-I and RMG-I-H cells were tested by quantitative Real-Time RT-PCR. The mRNA level of α1, 2-FT was significantly increased, but the mRNA level of CD44 was almost the same in RMG-1-hFUT cells and RMG-1 cells. (**P < 0.01, * P > 0.05).
Mentions: The 2-ΔΔCT value of mRNA level of CD44 in RMG-I-H cells is 79% of that in RMG-I cells, which had no significant difference (P > 0.05), whereas the mRNA level of α1,2-FT in RMG-I-H cells was increased by 3.07 times of that in RMG-I cells detected by Real-time PCR (P < 0.01). (Figure 4).

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