Lewis y 抗原对人卵巢癌细胞系 RMG-I 中 CD44 介导的黏附和铺展的增强作用。
Enhancive effects of Lewis y antigen on CD44-mediated adhesion and spreading of human ovarian cancer cell line RMG-I.
机构信息
Department of Obstetrics and Gynecology, Shengjing Hospital Affiliated to China Medical University, Shenyang, 110004, P R of China.
出版信息
J Exp Clin Cancer Res. 2011 Feb 7;30(1):15. doi: 10.1186/1756-9966-30-15.
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.
背景
本研究旨在探讨细胞黏附分子 CD44 与 Lewis y 抗原之间的分子结构关系，并确定 Lewis y 抗原对卵巢癌细胞系 RMG-I 及其高表达细胞系 RMG-I-H 中 CD44 介导的黏附和铺展的影响。
方法
采用免疫细胞化学法检测 Lewis y 单克隆抗体处理前后 RMG-I 和 RMG-I-H 细胞中 CD44 的表达；采用 Western blot 法检测 Lewis y 抗原和 CD44 的表达；采用免疫沉淀和共聚焦激光扫描显微镜法确定 Lewis y 抗原与 CD44 之间的结构关系；观察 RMG-I 和 RMG-I-H 细胞在透明质酸（HA）上的黏附和铺展；采用实时 RT-PCR 法检测 RMG-I 和 RMG-I-H 细胞中 CD44mRNA 的表达。
结果
免疫细胞化学结果显示，RMG-I-H 细胞中 CD44 的表达明显高于 RMG-I 细胞（P＜0.01），Lewis y 单克隆抗体处理后两种细胞系中 CD44 的表达均明显降低（均 P＜0.01）。Western blot 证实 RMG-I-H 细胞中 CD44 的含量是 RMG-I 细胞的 1.46 倍。共免疫沉淀证实 Lewis y 抗原和 CD44 共定位。RMG-I-H 细胞中 CD44 和 Lewis y 抗原的共表达是 RMG-I 细胞的 2.24 倍。RMG-I-H 细胞在 HA 上的黏附和铺展明显增强，与 RMG-I 细胞相比差异有统计学意义（P＜0.01），Lewis y 单克隆抗体可抑制其增强作用（P＜0.01）。两种细胞系中 CD44 的 mRNA 水平相似（P＞0.05）。
结论
Lewis y 抗原增强卵巢癌细胞中 CD44 介导的黏附和铺展。
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