Screening And Confirmation The Different Gene From Human Breast Cancer Cell MDA-MB-231 And T47D After Genistein And Endogenous Estradiol Double Acting

Objective:Breast cancer is the highest incidence of malignant tumors of women,seriously affecting the quality of human-s life. Now,it is known that the incidence of breast cancer is the result of many abnormal gene expression,Looking for the process of breast cancer gene expression levels change targets and prevent tumor development cancer research has become urgent problem.A large number of epidemiological data shows that the incidence of tumor initiation and development process, all ingredients on food and can be adjusted, the genistein as an important component of an active ingredient from soy foods diet, which enables falling the incidence of breast cancer, received extensive attention. Which is mainly produce estrogen and anti-estrogen effect manifests its anti-cancer activity. As estrogenic hormones in the human body is the effect of its key points together on breast cancer to determine prevention and control of breast cancer. In addition, different types of estrogen receptors, there is a big difference between the biological effects of wood dye flavonoids.In this study, using the advantages of high-throughput gene chip to Screen and confirmate the different gene from human breast cancer cell MDA-MB-231 and T47D after genistein and endogenous estradiol double acting.Getting the different key genes and signaling pathways from different estrogen receptors in breast cancer cells.analysis the role of the differentially expressed genes path and explore the role of the interaction between genes and gene pathways suspicious gene cluster, find genistein inhibition the role of the molecular mechanisms of breast cancer and inhibition of differences under different estrogen receptor in breast cancer cells in their breast cancer prevention, treatment application on scientific evidence.Methods:Normalizing the microarray data which from estradiol and genistein combined effect in human breast cancer cell line MDA-MB-231 and T47D after treatment, according to the fold difference (FC<0.5 or FC>2) and ANOVA screening differentially expressed genes, FC>2 means that the upregulated genes, FC<0.5 means that the down-regulated gene expression. By DAVID (The Database for Annotation, Visualization and Integrated Discovery) online analysis tools to this group of differentially expressed genes and gene annotation enrichment analysis of gene function, screening associated with tumor progression of functional class, the difference in the selection of these functional classes and a plurality of multiple genes involved in higher functions like real-time quantitative PCR validation. Results:1、After using different concentrations of genistein and endogenous estradiol together on breast cancer cell line MDA-MB-231 and T47D, screening the microarray data for each treatment group differences in genetic, selection criteria is P<0.01, FC (Fold Chang)≥2 or FC<0.5. (1) When the two breast cancer cells have not joined genistein, just added to the corresponding estrogen test was 80,800 pmol/ L according to different menopause when estrogen levels in the body. In accordance with the selection criteria, breast cancer cell line MDA-MB-231 with a total of 134 genes were significantly changed, which raised 98, down 36; breast cancer cells T47D with a total of 122 genes significantly changed, which raised 85, down 37. (2) When the two breast cancer cells were added genistein concentration of 20 μ mol/L, according to premenopausal and postmenopausal estrogen levels in the body,the different concentrations of added estrogen test substance is 0,80,800 pmol/L. In accordance with the selection criteria, breast cancer cell line MDA-MB-231 in a total of 395 genes significantly changed, which raised 229, down 166; breast cancer cells T47D in a total of 503 genes significantly changed, which raised 313, down 190. (3) When the concentration of estrogen in two breast cancer cells menopause (80Pmol/ L) level, added genistein concentration of 20 μ mol/L,can obtaine the influence of genistein on the two levels of estrogen after menopause breast cancer cells result. In accordance with the selection criteria, breast cancer cell line MDA-MB-231 in a total of 151 genes significantly changed, which raised 57, down 94; breast cancer cells T47D in a total of 136 genes significantly changed, which raised 50, down 86. (4) When the two breast cancer cell estrogen hormone levels in premenopausal is (800Pmol/L) level, added genistein concentration of 20 μ mol/L,can obtaine the influence of genistein on the two levels of estrogen in premenopausal breast cancer cells result. In accordance with the selection criteria, breast cancer cell line MDA-MB-231 in a total of 56 genes were significantly changed, which raised 29, down 27; breast cancer cells T47D in a total of 80 genes were significantly changed, which raised 39, down 41.2、 In test substance E20 pmol/L, Gen 0μmol/L as control, when the test substance E2 of 0 pmol/L, Gen 20μmol/as test group, by real-time quantitative PCR experiments, screened from the experimental group out 6 gene expression levels BRCA2, CDKNIA, ABAT, FAS, TP53TG3, SDCBP and other than the control group significantly increased, TNF gene expression level was significantly reduced, consistent with the microarray results; EGFR gene expression level was significantly reduced, microarray results are inconsistent.Conclusions:Gen have played the protective effects to ERα-negative breast cancer cell line MDA-MB-231 either premenopausal or postmenopausal; Gen proliferate and inhibite theT47D growth when estrogen levels before menopause, Gen promote the T47D growth when estrogen levels after menopause.Gen impact on breast cancer cells in addition to associated with estrogen levels, but also in related to the type the ER of breast cancer cells.Gen mainly have some impact on the level of phosphorylation and transcriptional level in the ER signaling pathways; the specific performance of the role of estrogen receptor ER-α’s, Gen preventing ER-a 537th tyrosine residues groups inhibits phosphorylation of tyrosine kinase activity, so ER dimerization can not be formed, block its signal transduction.