Effects Of HPs And HS On The Invasion And Metastasis Of Breast Caner Cells

Aims：1. To investigate the effects of HS synthesis of MCF-10A and heparins (HPs) ofdifferent molecular weight on proliferation of breast cancer cells.2. To investigate the effects of HS and HPs on the metastasis and invasion of breastcancer cells.3. To explore the effects of HPs and HS on the expression of MMPs and HSPE.Methods:1. The HS chain was extracted in MCF-10A cell culture medium. The MCF-7,MDA-MB-231and MDA-MB-435have been treated with different concentrations ofHS and HPs for24,48and72hours, then the cell viability was detected with MTTassay. The IC50of HS and HPs were calculated according to this experiment.2. Taking MCF-7, MDA-MB-231and MDA-MB-435as tool cells and investigating theeffects of HS and HPs on the invasion and metastasis of these cells. The metastaticactivity was observed by the wound healing assay.3. Taking MDA-MB-231and MDA-MB-435cells as tool cells and treating the cells byHS and HPs for24hours. The expressions of MMP-2and MMP-9were detected byWestern blotting and the HSPE activity was measured by the method of ELISA.Results:1. Anti-proliferative effects of HS and HPs on MCF-7, MDA-MB-231andMDA-MB-435cellsMTT assay showed that the proliferation of the breast cancer cells can be inhibitedby HS and HPs. The powerful order of the inhibition of the highest dose of HPs and HPon MCF-7cells was in turn HS, LMWH-B, LMWH-A and UFH. The comparison of thegroups treated by HS and LMWHs and those treated by UFH is significant (P<0.05).The powerful orders of the inhibition of the highest dose of HPs and HP onMDA-MB-231and MDA-MB-435cells were also in turn HS, LMWH-B, LMWH-Aand UFH. The comparisons of the groups treated by HS and LMWHs and those treatedby UFH were significant (P<0.05) as well. 2. Anti-invasive and anti-metastatic effects of HS and HPs on MCF-7,MDA-MB-231and MDA-MB-435cells2.1HS and HPs decreased the numbers of cells penetrated through the membrane intranswell invasion model, the comparisons with the control group were significant(P<0.01) and the decreasing-degree order of the invasive cells was in turn HS,LMWH-B, LMWH-A and UFH. The comparisons of the groups treated by HS andLMWHs and those treated by UFH were significant (P<0.05).2.2HS and HPs decreased the numbers of cells penetrated through the membrane inTranswell metastasis model, the comparisons with the control group were significant(P<0.01) and the decreasing-degree order of the invasive cells was in turn HS,LMWH-B, LMWH-A and UFH. The comparisons of the groups treated by HS andLMWHs and those treated by UFH were significant (P<0.05).2.3After treated by HS and HPs, the borderlines of the scarification were smooth andno cells were found in the scratch region at0h. The scarification healed in differentextend at24h and several cells metastasized into the scratch region. The scarification ofthe control group almost completely healed at48h, while the healing degree order of theHS and HPs groups was HS, LMWH-B, LMWH-A and UFH.3. Effects of HS and HPs on the expression of MMP-2and MMP-9in MCF-7andMDA-MB-435cellsBoth HS and HPs decreased the expression of MMP-2and MMP-9in MCF-7andMDA-MB-435cells, and the decreasing-degree order was HS, LMWH-B, LMWH-Aand UFH. While MMP-2and MMP-9scarcely expressed in weakly invasive andmetastatic MCF-7cells.4Effects of HS and HPs on the activity of HSPE in MCF-7and MDA-MB-435cellsBoth HS and HPs decreased the activity of MMP-2and MMP-9in MCF-7andMDA-MB-435cells, and the decreasing-degree order was HS, LMWH-B, LMWH-Aand UFH.Conclusion1. All of HS, LMWH and UFH may inhibit the proliferation, invasion and metastasis ofMCF-7, MDA-MB-231and MDA-MB-435breast cancer cells, the powerful order arein turn HS, LMWH-B, LMWH-A and UFH.2. The expressin of MMP-2and MMP-9as well as the activity of heparanase of MCF-7,MDA-MB-231and MDA-MB-435breast cancer cells all may be inhibited by HS,LMWH and UFH. The inhibitive powerful order was in turn HS, LMWH and UFH too.This corresponds with the suppression of the drugs on the invasion and mestastasis of MCF-7, MDA-MB-231and MDA-MB-435breast cancer cells.