摘要
目的采用乳腺癌和雌激素受体信号通路DNA oligo microarray功能基因芯片技术,观察金雀异黄素(genistein)对雌激素依赖性乳腺癌细胞T47D乳腺癌相关的基因表达及雌激素受体依赖性信号转导通路的影响。方法 0.001μmol/L雌二醇、10μmol/L金雀异黄素分别处理T47D细胞48 h,收集细胞并提取总mRNA,通过逆转录反应将mRNA标记,并与Oligo GEArray芯片杂交,芯片经计算机扫描图像采集得出差异表达基因。结果细胞周期蛋白(cyclin)基因Cyclin E1,CyclinE2与Cyclin A2表达均显著上调;ERα基因,雌激素诱导的相关基因PGR,TFF1/pS2,BCL2,ERBB2,C3,CCND1,TEF3,IGFBP2,IGFBP5,GATA3等也分别不同程度的上调,并采用实时荧光定量RT-PCR检测金雀异黄素可诱导pS2mRNA表达。结论金雀异黄素激活人乳腺癌T47D细胞ERα受体表达,通过周期蛋白过表达而调节细胞周期蛋白依赖性激酶(CDK)的活性,加速S期进程,促进细胞的增殖,并且雌激素诱导基因的上调显示了金雀异黄素的植物雌激素作用。
Objective To observe the influences of genistein on the gene expression of estrogen （E2 ）- dependent breast cancer T47D cells and estrogen receptor-dependent signal transduetion pathway by applying DNA oligo microarray technique. Methods T47D cells were respectively processed with 0.001 μmol/L E2 and 10 μmol/L genistein for 48 hours. The total mRNA was extracted after collecting cells, labeled through reverse transcription reaction, and then hybridized with Oligo GEArray？ chips. Differential expression genes were collected from the chips after computer scanning. Results The expressions of Cyclin El, Cyclin E2 and Cyclin A2 were all up-regulated significantly. The expressions of ERa, and E2-induced PGR, TFF1 （pS2 ）, BCL2, ERBB2, C3, CCND1, TEF3, IGFBP2, IGFBP5 and GATA3 were also up-regulated in varying degrees. The results of real-time fluorescence quantitative polymerase chain reaction （RT-PCR） showed that genistein induced the expression of pS2 mRNA. Conclusion Genistein can activate the expression of ERa receptor in T47D cells, regulate cyclin dependent kinase（CDK） activity to accelerate S phase, improve the proliferation of T47D cells, and play a role of phytoestrogen through up-regulating Ei-induced genes.
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2013年第2期100-103,共4页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(No.307725849)
河北省自然科学基金资助项目(No.H2012405016)
