Desouki Mohamed Mokhtar, Rowan Brian G
Department of Biochemistry and Molecular Biology, Medical College of Ohio, Toledo, Ohio 43614, USA.
Clin Cancer Res. 2004 Jan 15;10(2):546-55. doi: 10.1158/1078-0432.ccr-0661-03.
The purpose of this research was to determine whether a correlation exists between the levels of activated mitogen-activated protein kinase (MAPK) and Src kinases and the progression from normal to malignant endometrium.
We measured total and phosphorylated levels for extracellular signal-regulated kinase 1/2, p38, stress-activated protein kinase/c-Jun NH(2)-terminal kinase, and Src kinases from 33 frozen endometrial adenocarcinomas and 38 benign endometrial specimens by quantitation of signals from Western blots using antibodies against these kinases.
Elevated phospho-extracellular signal-regulated kinase 1/2 (150 +/- 40 versus 46 +/- 7; P = 0.03), phospho-Src (28 +/- 5 versus 4 +/- 1), and phospho-p38 (131 +/- 16 versus 27 +/- 7; P < 0.001) was detected in benign versus malignant endometrium when the Western blot signal of activated kinase was normalized to total kinase levels and beta actin. A modest increase in active c-Jun NH(2)-terminal kinase was detected in carcinoma versus benign specimens (51 +/- 13 versus 43 +/- 10; P = 0.8). Expression of total kinases (normalized to beta-actin) was higher in carcinoma versus benign specimens, respectively (extracellular signal-regulated kinase 1/2, 9 +/- 2 versus 0.7 +/- 0.1; Src, 7 +/- 2 versus 0.4 +/- 0.1; stress-activated protein kinase c-Jun NH(2)-terminal kinase, 2 +/- 0.4 versus 0.2 +/- 0.02; P < 0.001; and p38, 1 +/- 0.2 versus 0.4 +/- 0.1; P < 0.01). Immunohistochemistry for active and total Src kinases and MAPKs detected positive staining in epithelial and stroma cells.
These data demonstrated that, in contrast with breast cancer, the progression from normal to malignant endometrium is not associated with activation of MAPK and Src kinases. Elevation of these active kinases in benign endometrium may contribute to endometrial resistance to the antiestrogen action of tamoxifen.
本研究旨在确定活化的丝裂原活化蛋白激酶（MAPK）和Src激酶水平与子宫内膜从正常到恶性进展之间是否存在相关性。
我们通过使用针对这些激酶的抗体对蛋白质印迹信号进行定量，测量了33例冷冻子宫内膜腺癌和38例良性子宫内膜标本中细胞外信号调节激酶1/2、p38、应激激活蛋白激酶/c-Jun氨基末端激酶和Src激酶的总水平和磷酸化水平。
当将活化激酶的蛋白质印迹信号标准化为总激酶水平和β-肌动蛋白时，在良性与恶性子宫内膜中检测到磷酸化细胞外信号调节激酶1/2升高（150±40对46±7；P = 0.03）、磷酸化Src升高（28±5对4±1）以及磷酸化p38升高（131±16对27±7；P < 0.001）。在癌组织与良性标本中检测到活性c-Jun氨基末端激酶有适度增加（51±13对43±10；P = 0.8）。癌组织与良性标本中总激酶（标准化为β-肌动蛋白）的表达分别更高（细胞外信号调节激酶1/2，9±2对0.7±0.1；Src，7±2对0.4±0.1；应激激活蛋白激酶c-Jun氨基末端激酶，2±0.4对0.2±0.02；P < 0.001；以及p38，1±0.2对0.4±0.1；P < 0.01）。活性和总Src激酶以及MAPK的免疫组织化学检测在上皮细胞和基质细胞中呈阳性染色。
这些数据表明，与乳腺癌不同，子宫内膜从正常到恶性的进展与MAPK和Src激酶的激活无关。良性子宫内膜中这些活性激酶的升高可能导致子宫内膜对他莫昔芬抗雌激素作用产生抵抗。
