Kayed Hany, Kleeff Jörg, Kolb Armin, Ketterer Knut, Keleg Shereen, Felix Klaus, Giese Thomas, Penzel Roland, Zentgraf Hanswalter, Büchler Markus W, Korc Murray, Friess Helmut
Department of General Surgery, University of Heidelberg, Heidelberg, Germany.
Int J Cancer. 2006 Jan 1;118(1):43-54. doi: 10.1002/ijc.21257.
The expression and localization of FXYD domain containing ion transport regulator 3 (FXYD3), a transmembrane protein that acts as a chloride channel or chloride channel regulator, was analyzed in pancreatic tissues derived from donors and patients suffering from chronic pancreatitis (CP) or pancreatic ductal adenocarcinoma (PDAC) as well as in pancreatic cancer cells using QRT-PCR, laser-capture microdissection and microarray analysis, in situ hybridization and immunohistochemistry. FXYD3 antisense expressing T3M4 pancreatic cancer cells were generated and compared to control cells using anchorage-dependent and independent growth assays, and xenotransplantation into nude mice. FXYD3 mRNA levels were 3.4-fold increased in PDAC tissues compared to donor specimens (p = 0.006), and 3.9-fold increased in microdissected cancer cells compared to normal pancreatic ductal cells (p = 0.02). FXYD3 was localized in the tubular complexes and PanIN lesions of both CP and PDAC, as well as in pancreatic cancer cells. Downregulation of FXYD3 by stable antisense transfection increased significantly the doubling time of T3M4 pancreatic cancer cells from 44 +/- 2 hr to 55 +/- 12 hr (p = 0.02). Nude mice transplanted with antisense transfected cells displayed a significant increase in tumor doubling time from 3.3 days +/- 1.0 to 4.3 days +/- 0.43 (p = 0.058). Anchorage-independent growth and sensitivity to 5-fluorouracil, gemcitabine and cisplatin as well as to MgCl(2) were not dependent on the level of FXYD3 expression. In conclusion, overexpression of FXYD3 in pancreatic cancer may contribute to the proliferative activity of this malignancy.
含FXYD结构域的离子转运调节蛋白3（FXYD3）是一种跨膜蛋白，可作为氯离子通道或氯离子通道调节剂。采用实时定量聚合酶链反应（QRT-PCR）、激光捕获显微切割及微阵列分析、原位杂交和免疫组织化学方法，分析了FXYD3在慢性胰腺炎（CP）或胰腺导管腺癌（PDAC）供体及患者的胰腺组织以及胰腺癌细胞中的表达和定位。构建了FXYD3反义表达的T3M4胰腺癌细胞，并通过贴壁依赖性和非依赖性生长试验以及裸鼠异种移植实验与对照细胞进行比较。与供体标本相比，PDAC组织中FXYD3 mRNA水平升高3.4倍（p = 0.006），与正常胰腺导管细胞相比，显微切割得到的癌细胞中FXYD3 mRNA水平升高3.9倍（p = 0.02）。FXYD3定位于CP和PDAC的管状复合体和胰腺上皮内瘤变（PanIN）病变以及胰腺癌细胞中。通过稳定的反义转染下调FXYD3可使T3M4胰腺癌细胞的倍增时间从44±2小时显著增加至55±12小时（p = 0.02）。移植了反义转染细胞的裸鼠肿瘤倍增时间从3.3天±1.0显著增加至4.3天±0.43（p = 0.058）。非贴壁生长以及对5-氟尿嘧啶、吉西他滨、顺铂和MgCl₂的敏感性不依赖于FXYD3的表达水平。总之，FXYD3在胰腺癌中的过表达可能有助于这种恶性肿瘤的增殖活性。
