Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Harbin Medical University, Harbin, 150081, China.
Mol Cell Biochem. 2012 Jan;359(1-2):235-43. doi: 10.1007/s11010-011-1018-8. Epub 2011 Aug 12.
A disintegrin and metalloprotease 17 (ADAM17) is a transmembrane protein that can cleave membrane anchored proteins to release soluble factors and regulate important biological phenomena in cancers. In the present study, we evaluated the effects of ADAM17 on the proliferation and on the cell cycle distribution of human prostate cancer cells. Experiments were also performed to gain insights into the possible mechanism of action of ADAM17. We used over-expression and RNAi strategy to investigate the function of ADAM17 in human prostate cancer cells. Changes in rate of proliferation and cell cycle profile were measured by growth curve, Cell Counting Kit-8 (CCK-8) assay, bromodeoxyuridine (BrdU) incorporation assay and cell cycle analysis. In addition, changes in expression of associated genes and proteins were studied by semiquantitative RT-PCR, western blotting and ELISA analysis. Ectopic over-expression of ADAM17 resulted in increased cell proliferation. We also showed that ADAM17 promoted G1 to S phase transition concomitantly with upregulation of cyclin E, CDK2 and downregulation of p21 and p27 proteins. ADAM17 over-expression cells showed that more TGF-α released to the supernatant and activated the EGFR/PI3K/AKT pathway. Conversely, silencing ADAM17 led to the opposite effect. Both siRNAs knockdown of ADAM17 and blocking the EGFR/PI3K/AKT pathway using specific inhibitor caused downregulation of cyclin E, CDK2, and upregulation of p21 and p27 in prostate cancer cells. Collectively, this study demonstrates that over-expression of ADAM17 might target cyclin E, CDK2, p21, and p27 to promote prostate cancer cell proliferation through activation of the EGFR/PI3K/AKT pathway.
解整合素金属蛋白酶 17（ADAM17）是一种跨膜蛋白，能够切割膜锚定蛋白以释放可溶性因子，并调节癌症中的重要生物学现象。在本研究中，我们评估了 ADAM17 对人前列腺癌细胞增殖和细胞周期分布的影响。还进行了实验以深入了解 ADAM17 的可能作用机制。我们使用过表达和 RNAi 策略来研究 ADAM17 在人前列腺癌细胞中的功能。通过生长曲线、Cell Counting Kit-8（CCK-8）测定、溴脱氧尿苷（BrdU）掺入测定和细胞周期分析来测量增殖率和细胞周期谱的变化。此外，通过半定量 RT-PCR、Western blot 和 ELISA 分析研究相关基因和蛋白的表达变化。ADAM17 的异位过表达导致细胞增殖增加。我们还表明，ADAM17 促进了 G1 到 S 期的转变，同时上调了细胞周期蛋白 E、CDK2，并下调了 p21 和 p27 蛋白。ADAM17 过表达细胞显示更多的 TGF-α释放到上清液中，并激活了 EGFR/PI3K/AKT 途径。相反，沉默 ADAM17 导致相反的效果。使用特异性抑制剂沉默 ADAM17 和阻断 EGFR/PI3K/AKT 途径都导致前列腺癌细胞中细胞周期蛋白 E、CDK2 的下调和 p21、p27 的上调。总之，这项研究表明，ADAM17 的过表达可能通过激活 EGFR/PI3K/AKT 途径靶向细胞周期蛋白 E、CDK2、p21 和 p27，促进前列腺癌细胞增殖。
