Wilkins Heather R, Doucet Kristin, Duke Victoria, Morra Amber, Johnson Nicole
Department of Natural Sciences, Assumption College, 500 Salisbury Street, Worcester, MA 01609, USA.
Tumour Biol. 2010 Jan;31(1):16-22. doi: 10.1007/s13277-009-0003-2. Epub 2009 Dec 18.
The proto-oncogene c-myb is overexpressed in human colon cancer cells. c-myb is known to be affected by estrogen in some breast cancers and leukemias. However, the mechanism of c-myb regulation via estrogen in colon cancer requires further investigation. Human COLO-205 colon cancer cells were cultured and treated with beta-estradiol for 24 h. Apoptosis was quantified using acridine orange/propidium iodide labeling and confirmed with DNA fragmentation gel electrophoresis. Expression of c-myb protein was assessed via SDS-PAGE and immunoblotting and RT-PCR was used to quantify bcl-2 RNA. Protein and RNA expression levels were also assayed after c-myb siRNA treatment for 24 h. We demonstrate an increase in apoptosis after 24 h of beta-estradiol treatment of human COLO-205 colon cancer cells. Estrogen treatment also decreases c-myb protein levels as well as expression of its transcriptional target bcl-2. Suppression of c-myb protein also results in increased apoptosis and decreases bcl-2 expression. These results indicate that estrogen has a protective effect from sustained colon cancer cell growth at least partly through suppression of c-myb and bcl-2.
原癌基因c-myb在人结肠癌细胞中过表达。已知c-myb在某些乳腺癌和白血病中受雌激素影响。然而,在结肠癌中通过雌激素调节c-myb的机制需要进一步研究。培养人COLO-205结肠癌细胞并用β-雌二醇处理24小时。使用吖啶橙/碘化丙啶标记定量凋亡,并用DNA片段凝胶电泳进行确认。通过SDS-PAGE和免疫印迹评估c-myb蛋白的表达,并用RT-PCR定量bcl-2 RNA。在c-myb siRNA处理24小时后也检测蛋白质和RNA表达水平。我们证明,人COLO-205结肠癌细胞经β-雌二醇处理24小时后凋亡增加。雌激素处理还降低了c-myb蛋白水平及其转录靶标bcl-2的表达。c-myb蛋白的抑制也导致凋亡增加并降低bcl-2表达。这些结果表明,雌激素至少部分地通过抑制c-myb和bcl-2对持续的结肠癌细胞生长具有保护作用。