RESUMO
<p><b>OBJECTIVE</b>To observe the effect of grape seed extract (GSE) on breast cancer cell MCF-7 about the of proliferation and the gene expression of survivin, and to investigate the molecular biological mechanism of inhibition by GSE.</p><p><b>METHOD</b>MTT was used to measure the role of proliferated inhibition of GSE at the dosage of 40, 80, 120, 160, 200 mg x L(-1) with different time. To calculate the IC50 of GSE and to select the suitable concentration and treatment time. The change of cell cycle was detected by flow cytometry. mRNA expression of Survivin was observed by RT-PCR. Luciferase kit was used to observe the change of core promoter of survivin which was inserted in the flourescence report vector. And further to analyse the bind side of transcription factor on the sequence of core promoter of survivin was further analysed by using the microsoft of bioinformatics.</p><p><b>RESULT</b>GSE could inhibit the proliferation of breast cancer cell MCF-7 with time and dosage-dependent (P < 0.05). GSE could arrest the cell cycle in S periods (P < 0.05) and also inhibit the mRNA expression of survivin effectively (P < 0.01); GSE could down-regulate the activity of core promoter of survivin significantly (P < 0.01).</p><p><b>CONCLUSION</b>GSE can inhibit the proliferation of breast cancer cell MCF-7 through arresting the cell cycle in S periods. The mechanism may be that GSE regulate the activity of transcription factor which are related to the activity of core promoter of survivin and decrease the gene expression of survivin.</p>
Assuntos
Feminino , Humanos , Antineoplásicos Fitogênicos , Farmacologia , Neoplasias da Mama , Genética , Metabolismo , Patologia , Linhagem Celular Tumoral , Proliferação de Células , Clonagem Molecular , Regulação para Baixo , Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos , Proteínas Inibidoras de Apoptose , Proteínas Associadas aos Microtúbulos , Genética , Metabolismo , Transfecção , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Objective Investigate the effect of TrKA variation on the expression of neucleoprotein NF-?B P65 and apoptosis.Methods To construct the expression vector of TrKA small interfering RNA,the recombinant was transfected into MCF-7 cells.the stable cell line expressing TrKA small interfering RNA were selected by G418.The mRNA and protein of TrKA were tested by real-time PCR,Western-blot and Immunohistochemistry.The change of neucleoprotein NF-?B P65 was detected by WB,Flow cytometry was used to observe the cell apoptosis.ResultsThe expression vector of TrKA-siRNA was successfully constructed.The mRNA and protein of TrKA were decreased by 74.7% and 80.5% respectively(P