Inhibition of MUC4 expression suppresses pancreatic tumor cell growth and metastasis
Inhibition of MUC4 expression suppresses pancreatic tumor cell growth and metastasis
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DOI：
10.1158/0008-5472.can-03-2636
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发表时间：
2004-01-15
期刊：
影响因子：
11.2
通讯作者：
Batra, SK
中科院分区：
文献类型：
--
作者：
Singh, AP;Moniaux, N;Batra, SK
MUC 4粘蛋白是一种高分子量的膜结合糖蛋白。它在胰腺肿瘤和肿瘤细胞系中异常表达，而在正常胰腺中检测不到表达。在胰腺上皮内瘤变中也观察到MUC 4表达的进行性增加，表明其与疾病发展相关。在这里，我们研究了在表达高水平MUC 4的侵袭性和高转移性胰腺肿瘤细胞系CD 18/HPAF中MUC 4表达沉默的后果。通过表达反义MUC 4 RNA的质粒构建体的稳定整合下调MUC 4的表达。通过Western印迹和免疫荧光分析证实，MUC 4表达的降低导致与亲本、空载体（ZEO）和正义转染（ES6）对照细胞相比，反义MUC 4转染（EIAS 19）细胞的生长和克隆形成能力降低。此外，当原位移植到免疫缺陷小鼠中时，EIAS 19细胞显示出肿瘤生长和转移特性的显著降低。在运动，粘附，和聚集的体外生物学测定表明，3倍的减少与对照细胞相比，EIAS 19细胞的运动，而这些细胞粘附更多，并表现出细胞聚集的增加。有趣的是，MUC 4下调也与其假定的相互作用伴侣HER 2/neu在反义MUC 4转染细胞中的表达减少相关。总之，本研究首次证明了MUC 4粘蛋白与转移性胰腺癌表型的直接相关性，并为MUC 4在肿瘤细胞生长和行为特性改变中的功能作用提供了实验证据。
The MUC4 mucin is a high molecular weight membrane-bound glycoprotein. It is aberrantly expressed in pancreatic tumors and tumor cell lines with no detectable expression in the normal pancreas. A progressive increase of MUC4 expression has also been observed in pancreatic intra-epithelial neoplasia, suggesting its association with disease development. Here, we investigated the consequences of silencing MUC4 expression in an aggressive and highly metastatic pancreatic tumor cell line CD18/HPAF that expresses high levels of MUC4. The expression of MUC4 was down-regulated by the stable integration of a plasmid-construct expressing antisense-MUC4 RNA. A decrease in MUC4 expression, confirmed by Western blot and immunofluorescence analyses, resulted in diminished growth and clonogenic ability of antisense-MUC4-transfected (EIAS19) cells compared with parental, empty vector (ZEO) and sense transfected (ES6) control cells. In addition, EIAS19 cells displayed a significant decrease in tumor growth and metastatic properties when transplanted orthotopically into the immunodeficient mice. In vitro biological assays for motility, adhesion, and aggregation demonstrated a 3-fold decrease in motility of EIAS19 cells compared with control cells, whereas these cells adhered more and showed an increase in cellular aggregation. Interestingly, MUC4 down-regulation also correlated with the reduced expression of its putative interacting partner, HER2/neu, in antisense-MUC4-transfected cells. In conclusion, the present work demonstrates, for the first time, a direct association of the MUC4 mucin with the metastatic pancreatic cancer phenotype and provides experimental evidence for a functional role of MUC4 in altered growth and behavioral properties of the tumor cell.