Relationships among Anti-mastocarcinoma Activity of Emodin and ERα Pat Hway and Immune Function
|School||Liaoning Normal University|
|Keywords||Emodin Breast cancer ERα Signaling pathway immunoregulation|
Breast cancer is an estrogen-dependent tumors,There is accumulating evidenceimplicating that estrogen stimulation is an important virulence factor for breast cancer,itsbiological activity is regulated by estrogen receptor(ERα).ERα-mediated genomic signalingpathways and non-genomic signaling pathways play the estrogen proliferation andanti-apoptotic effects, abnormal activation of the signaling pathway is the leading cause ofbreast cancer.Currently, selective estrogen receptor modulator tamoxifen (tamoxifen) as therepresentative of the most representative of the anti-estrogen drugs, and its mechanism ismainly targeted intervention ERα signal pathway.But following the deepening of thetreatment in patients who appear with varying degrees of resistance, and most of the bodyappear to side effects，such as immune function, suppression and destruction.Therefore, thesearch the toxic side effects is small, safe and effective an antineoplastic drug become the thea research hotspot of the in recent years scholars from various countries.Therefore, findinglittle toxic and side effects, safe and effective antitumor drugs has become a research hotspotin recent years.In recent years, natural products has obvious characteristics of the antitumor activity andlow toxicity and subject to people’s attention.Natural anthraquinone compounds exist widelyin nature, it has many pharmacological effects, may play a antitumor role through differentmechanisms.At present, commonly used in clinical anticancer drugs doxorubicin andmitoxantrone basic nucleus is anthraquinone structure, suggesting that having a mothernucleus structure of the anthraquinone-based compounds have anti-tumor activity.Emodin is astrong anti-cancer activity of natural compounds of anthraquinone from rhubarb, Polygonumcuspidatum, aloe vera in separation, anti-tumor and immune regulation effect is the hotspot ofthe research.Due to the lack of in-depth study of emodin against breast cancer action andsignal mechanism, about the relationship of emodin against breast cancer and ERα signalingpathway, both at home and abroad is seldom reported, so this experiment of emodin in vitroagainst the relationship between breast cancer and ERα signaling pathway were studied, andthe detection of the immunomodulatory effects of emodin, aims to provide a new drug againstbreast cancer some theoretical basis for the development and application.The results are asfollows: 1.MTT assay was used to detect emodin in vitro against the proliferation of breast cancercells, experimental results show that the ERα-positive MCF-7cells and ERα-negativeMDA-MB-231cell proliferation was inhibited, but on the the ERα positive MCF-7cellproliferation is more obvious, by a significant dose and time dependent manner.2.MTT assay and flow cytometry detection of emodin on the E2mitogenic andanti-apoptotic role. The results show that the E2is able to significantly promote theproliferation of MCF-7cells in vitro and inhibit apoptosis, emodin effective inhibition ofE2-induced MCF-7cells in vitro and anti-apoptotic effect, the E2-induced MCF-7cell cyclearrested in G0/G1phase.3.Using Western blotting method and luciferase assay method, detection of the effect ofemodin on estrogen receptor ERα protein expression and transcriptional activity of MCF-7,the experimental results show, emodin had significant inhibitory effect on ERα proteinexpression and transcriptional activity, in a dose-dependent manne.4.By Western blotting detection of emodin on ERα target gene the protein CyclinD1andBcl-2expression.The results show that emodin expression had significant inhibitory effect onMCF-7cells of ERα gene protein Cyclin D1and Bcl-2.5.Using Western blotting method, detection of emodin E2fast MCF-7cells induced byMAPK/Erk1/2and PI3K/Akt signal pathways were detected. The results showed that emodincan inhibit the E2-induced Akt and MAPK protein phosphorylation, but more significantinhibition of protein activity. Emodin ERα non-genomic signaling pathways throughPI3K/Akt signaling pathways to achieve.6.The phagocytosis of neutral red assay and MTT colorimetric method, effect of emodinon nonspecific immune cells. The results show, emodin can improve the metabolic activity ofperitoneal macrophages and phagocytic ability, in a dose-dependent manner. At the same time,emodin can significantly increase the activity of NK cells, with the increase of drugconcentration, the activity of NK cells increased gradually. Emodin has a role in promotingthe non-specific immune cells.7.MTT assay was used to detect the impact of of emodin specific immune cells. Theresults showed that emodin mouse spleen T lymphocytes and B lymphocytes cytotoxic effects,no significant effect on promoting their proliferation.8.Hemolysis in vitro experiments, detection of emodin human erythrocytes. The resultsshowed that emodin various concentrations of hemolysis and erythrocyte agglutination,hemolysis rate <5%, indicating that emodin is a safe and non-toxic natural products.