BACKGROUND: Angiogenesis is an important prognostic indicator for malignant tumors. Breast cancer overexpressing oncogene HER-2/neu often denotes a poor prognosis. Many studies have demonstrated the antitumor effect of genistein against breast cancer.OBJECTIVE: To study the relationship between HER-2/neu expression and angiogenesis in breast cancer as well as the effect of genistein on the angiogenesis in HER-2/neu-overexpressing breast cancer.DESIGN: A randomized controlled observatory experiment with nude mice.SETTING: Department of nutrition and food hygiene of a military medical university.MATERIALS: Twenty specific pathogen-free(SPF) normal female BALB/c nude mice weighing (10 ± 2) g, aged 3 to 4 weeks, were purchased from the Experimental Animal Center of the Third Military Medical University.METHODS: This study was carried out in the Department of Nutrition and Food Hygiene, Third Military Medical University from June 2001 to March 2002. HER-2/neu-overexpressing breast cancer cell line MCF-7/HER-2 was generated by transfecting MCF-7 cells with human HER-2/neu cDNA. MCF-7/HER-2 and MCF-7 cells were inoculated in female BALB/c nude mice to establish tumor-bearing mouse models. Four weeks after the inoculation, the mice with MCF-7/HER-2 xenografts were randomly divided into control,genistein treatment, and anti-HER-2/neu antibody treatment groups to receive corresponding treatments every other day for two weeks, at the end of which the tumor volume, microvessel density(MVD) and vascular endothelial growth factor(VEGF) expression in the xenografts were measured.MAIN OUTCOME MEASURES: MVD and VEGF expression in the xenograft tumor. Secondary outcome measures: Identification of HER-2/neu-transfected from MCF-7-transfected cells and the tumor volume.RESULTS: The MVD was 16 ±6, 98 ±21, 56± 18, and 52 ± 19 in each visual field in the MCF-7 xenografts group, control group, genstein treatment group and anti-HER-2/neu antibody treatment group recpectively. MVD and VEGF expression in MCF-7/HER-2 xenografts were higher than that in MCF-7 xenografts, and was reduced after treatment with genistein or anti-HER-2/neu antibody. The changes of tumor volume in these xenografts were consistent with the changes of MVD and VEGF.CONCLUSION: HER-2/neu overexpression in breast cancer promotes angiogenesis, and genistein can inhibit angiogenesis and growth of HER-2/neu-overexpressing breast cancer to improve the prognosis.

Objective　To study the effect of glucagon-like peptide 2 (GLP-2) on mitogen-activated protein kinase (MAPK) activity in small intestinal epithelia in mice after radiation injury and its relation with the change of small intestinal epithelial proliferation. Methods　Mice were given a single dose of 8 Gy of total body 60Co gamma irradiation and then divided into GLP-2 and control groups. The activity of MAPK and proliferation rate in small intestinal epithelia were measured. Results　The activity of MAPK in small intestinal epithelia was higher in GLP-2-treated mice than in irradiated mice, and the proliferation rate in small intestinal epithelia significantly increased in GLP-2-treated mice. These two indices were of significantly positive correlated. Conclusion　GLP-2 can promote small intestinal epithelial proliferation in irradiated mice, and this may be related to activation of MAPK in small intestinal epithelia.

Objective　To investigate the effects of glucagon-like peptide 2 (GLP-2) on recovery of small intestinal epithelia from radiation injury in mice. Methods　Mice received a single 8 Gy dose of total body irradiation from 60Co gamma ray followed by treatment with GLP-2 or vehicle. DNA and protein content in small intestinal mucosa were measured, and small intestine was processed for histological examination with light microscope and scanning electron microscope. Results　Small intestinal mucosal DNA and protein content, villus height, and villus number significantly decreased in irradiated mice, partial villus tips were ulcerated. GLP-2 administration caused increase in DNA and protein content, villus height, and villus number as compared with irradiated control group. Meanwhile, the villus tips were lack of ulceration. Conclusion　GLP-2 can promote recovery of small intestinal epithelia from radiation injury in mice.

Objective To observe the effect of genistein,octylphenol,and their combination on proliferation of MCF-7 cells and explore the molecular mechanisms.Methods The MCF-7 cells were divided into four groups: control,5?10~(-5)mol/L genistein treated,8?10~(-6)mol/L octylphenol treated,genistein and octylphenol treated.The cell proliferation,cell cycle,phospho-L-tyrosine protein(PTPr),ER?,ER? mRNA and AIB1 mRNA expression of MCF-7 cells was observed by MTT test,flow cytometry,immunocytochemistry and RT-PCR.Results When MCF-7 cells were treated with 8?10~(-6) mol/L octylphenol or 5?10~(-5) mol/L genistein,or both for 72 h,the proliferation ratio was 12.98%,46.16% and 36.44% respectively;the percentage of MCF-7 cells at G_(2)/M were 12.98%,46.16% and 36.44% respectively;the apoptosis ratio of MCF-7 cells were 3.57%,11.41% and 8.24% respectively;the expression of PTPr was(62.84?9.80),(26.75?5.09),(39.15?7.83) respectively.Octylphenol increased the expression of AIB1 mRNA,but genistein and its combination with octylphenol inhibited the expression of ER? and decreased the expression of AIB1 mRNA in nucleus.Conclusion Octylphenol can promote the proliferation of MCF-7 cells,while genistein and its combination with octylphenol inhibit,which mechanism may be related to regulation of PTPr,AIB1 and ER expression.

Objective: To study the effects of octylpheno(lOP)and genistein(GEN)on the expressions of estrogen receptors(ER),amplified in breast cancer 1(AIB1) and proliferating cell nuclear antigen(PCNA) in rat breast cancer.Method:Female SD rats were randomly divided into control,model,GEN treated,OP treated,GEN and OP combined treated groups.RT-PCR and immunohistochemical methods were used to detect the expressions of AIB1,ER,PCNA on normal mammary gland and mammary cancer.Results: Compared with control mammary gland,AIB1 mRNA,ER mRNA,PCNA and ER expressions were up-regulated in mammary cancer.Compared with mammary cancer in model group,the level of AIB1mRNA,ER mRNA and ER expressions were significantly decreased in GEN treated group,while they were significantly increased in OP treated group and PCNA expression was significantly increased too.Compared with OP treated group,the level of AIB1mRNA,ER mRNA,ER and PCNA expressions were partly decreased in GEN and OP combined treated group.Conclusion: Octylphenol can up-regulate the levels of AIB1,ER and PCNA expressions in mammary cancer and may increase the incidence of 7,12-dinmethylbenz[a]anthracene(DMBA)-induced mammary cancer.Genistin or genistein combined with octylphenol can downregulate the levels of AIB1,ER and PCNA expressions in mammary cancer,and inhibit theiincidences.

Objective To investigate the mechanisms of genistein in the growth inhibition in the tumor angiogenesis. Methods The effects of genistein and vascular endothelial growth factor receptor (VEGFR) on the proliferation of ECV304 cells were observed by cell growth curves and flow cytometry. The level of the phosphorylation of VEGFR and the activity of protein tyrosine kinase (PTK) in ECV304 cells were detected by immunoprecipitation and kinase activity analysis. Results Vascular endothelial growth factor (VEGF) alone could facilitate the proliferation of ECV304 cells, up regulate the phosphorylation and PTK activity of VEGFR, but genistein alone could inhibit the growth of ECV304 cells, stop the cell cycle mainly at the phase of G 2/M, induce apoptosis, and down regulate the phosphorylation and PTK activity of VEGFR ( P

Objective: To study the effects of genistein on the expression of urokinase-type plasminogen activator (uPA) and the activity of protein tyrosine kinase (PTK) in MDA-MB-453 cells, and explore the molecular mechanism of anti-angiogenesis in HER-2/neu-overexpressing breast cancer by genistein. Methods:Western blot, immunoprecipitate, reverse transcription-polymerase chain reaction (RT-PCR) and kinase activity analysis technics were used to observe the expression of uPA and the protein phosphorylation of HER-2/neu receptor and the activity of protein tyrosine kinase (PTK) in MDA-MB-453 cells treated by genistein for 24, 48, 72 h. Results: The expression of uPA and the protein phosphorylation of HER-2/neu receptor and the activity of PTK were significantly decreased after treated with 5?10-5mol/L genistein, which had a time-dependence. Conclusion: Genistein could inhibit the activity of PTK and the protein phosphorylation of HER-2/neu receptor, and down-regulate the exprssion of uPA at transcription and translation levels in breast cancer cells. This might be a part of molecular mechanism of genistein anti-angiogenesis in HER-2/neu-overexpressing breast cancer.

Objective To explore the inhibitory effect of genistein on oncogene HER-2/neu-overexpressing breast cancer xenograft tumors and its relationship with urokinase type plasminogen activator (uPA) expression. Methods HER-2/neu-overexpressing MCF-7 human breast cancer cells (MCF-7/HER-2) were generated by transfecting the HER-2/neu negative MCF-7 cells with HER-2/neu cDNA. MCF-7/HER-2 and MCF-7 xenograft models were established in BALB/c nude mice, and the nude mice with MCF-7/HER-2 xenograft were randomly divided into three groups: control group, genistein-treated group, and HER-2/neu antibody-treated group. The growth ability of xenograft tumors was analyzed by immunohistochemical staining with Ki-67 antibody, and uPA expression of xenograft tumors was analyzed by Western blotting. Results The growth ability and uPA expression were significantly higher in MCF-7/HER-2 xenograft tumors than those in MCF-7 xenograft tumors. MCF-7/HER-2 xenograft tumors treated by genistein or HER-2/neu antibody showed lower growth ability and uPA expression as compared with MCF-7/HER-2 xenograft tumors. Conclusion Genistein can inhibit the growth ability of HER-2/neu-overexpressing breast cancer xenograft tumors, which is associated with the uPA expression.

Objective　To investigate the expression of TGF-β and TGF-β receptor in human breast cancer cell Bcap-37 inhibited by soybean isoflavones. Methods　mRNA and protein of TGF-β1、TGF-βRⅠ in Bcap-37 cells were examined with reverse transcription ploymerase chain reaction(RT-PCR) and immunohistochemistry after cells were treated with daidein or genistein for 1-4 d.The expression of TGF-β1 and TGF-β2 was determined with TGF-β resistance test. Results　The TGF-β1, TGF-β2 and TGF-β recepor increased in Bcap-37 cells at a concentration of 3×10-5 mol/L of genistein. No changes was found when treated with daidzein. Conclusion　Genistein may inhibit the proliferation of Bcap-37 cells and accompany with increasing expression of TGF-β1, TGF-β2 and TGF-β receptor.