Study On Safflower Polysaccharide About Inhibiting The Cell Proliferation And Influencing The Metastasizing Ability Of Human Breast Cancer Cell Line Mcf-7

Objective:To investigate nm23-H1, c-erbB-2and MMP-9protein expression in breast cancer tissue and its relationship with the relevant indicators and transfer; to study the influence of safflower polysaccharides (SPS), epirubicin (EPI) on the proliferation and apoptosis of the human breast cancer cells MCF-7, and observe the changes of the apoptosis and metastasis related genes of BAX, Bcl-2and MMP-9, nm23-H1mRNA and protein expression, to confirm using Transwell chamber assay that safflower polysaccharide, epirubicin can suppression the metastatic ability of breast cancer cells. And make sure that Chinese medicine safflower polysaccharide has the ability of anti-MCF-7cells by certain mechanism.Methods:1. Random collecting120cases of breast cancer tissue samples and77cases of normal breast tissue samples. We try to use SP immunohistochemistry method to detect the expression of nm23-H1, c-erbB-2and MMP-9in breast cancer tissue and normal breast tissue. To combine with clinical and pathological data, and analysis of the three exppression with age, clinical stage, lymph node metastasis the relationship of the clinical and pathological features.2. To observe the proliferation inhibition and apoptosis of human breast cancer MCF-7cell effected by safflower polysaccharide or epirubicin by using MTT assay and flow cytometry (FCM) analysis.3.To detect the expression levels of BAX and Bcl-2and MMP-9, nm23-Hl mRNA in MCF-7cell by using real-time PCR; to detect the expression levels of BAX and Bcl-2and MMP-9, nm23-H1mRNA in MCF-7cell by using western blot.4. Transwell chamber experiment was used to investigate effects on the metastatic ability of breast cancer cell line MCF-7by giving different concentrations of safflower polysaccharide or epirubicin.Results:1. Nm23-H1, c-erbB-2and MMP-9were highly expressed in breast cancer; only the c-erbB-2level related with age; expression of nm23-H1protein was high in the early stage of breast cancer and breast cancer with no lymph node metastasis or distant metastasis; however, expression of c-erbB-2and MMP-9were high in the late stage, lymph node metastasis and distant metastasis of breast cancer.2. Safflower polysaccharide and epirubicin can both inhibit the proliferation of breast cancer MCF-7cell. The depressant effect of Epirubicin on MCF-7cells increased with the concentration of Epirubicin, and decreased with time; the inhibition of safflower polysaccharide on MCF-7cells increased with time progresses, and in a certain concentration, as the concentration increased, the enhanced role increased, till the concentration achieved or larger than the certain concentration, and then the inhibition effect decreased.3. Be treated wiht SPS or EPI for24hours, flow cytometry was applied. Results showed that the early apoptosis rate, cell necrosis and late apoptosis rate of MCF-7cells were significantly increased, showing a significant concentration-dependent.4. Real-time PCR and western blot results showed that, treated with SPS and EPI, the expression of Bcl-2and MMP-9(both in RNA and protein) were decreased, and the level of BAX and nm23-H1mRNA were increased.5. Both SPS and EPI had the ability to inhibit metabasis of breast cancer MCF-7cells.Conclusion:1. High expression of c-erbB-2and MMP-9and low expression of nm23-H1were closely related to invasion and metastasis in breast cancer. Testing these index can be helpful in early diagnosis of metastasis and recurrence of breast cancer, providing evidence for estimating the outcome, and providing theoretical basis on anticancer targets of reference2. Inhibition effect of Traditional Chinese medicine SPS and EPI on breast cancer cell MCF-7cells was significant different. The inhibition of safflower polysaccharide on MCF-7cells increased with time progresses only in a certain concentration.3. Both SPS and EPI induced human breast MCF-7apoptosis and necrosis in a dose dependent; the effect of apoptosis and necrosis by SPS is weaker than that by EPI. 4. Both SPS and EPI can induce Bcl-2and MMP-9gene lowly express, and induce BAX and nm23-H1gene highly express.5. SPS and EPI both can inhibit the metastatic potentia of breast cancer MCF-7cells, with no significant difference.