The Study On The Proliferation Effect Of Breast Cancer Cells Conditioned Medium On Osteoblasts And Their Mechanisms

Objective: To study the effect on the proliferation of two different types of breast cancer cell conditioned medium on osteoblast,and to investigate preliminary whether the expression of protein and m RNA by regulating Notch signaling pathway in Notch1,Jagged1,Hes1 influence osteoblast proliferation activity,for the clinical treatment of breast cancer bone metastasis to provide a new potential therapeutic target.Methods:1.The effects on the proliferation were detected two different types of breast cancer cells(MDA-MB-231,MCF-7)conditioned medium(CM)with different concentrations(0,20%,40%,60%,80%,100%)and different time(0,24,48,72,96h)into a dose-dependent and time-dependent effect inhibited of osteoblast for by MTT;After interference by DAPT(Notch inhibitor)effects on osteoblasts;Flow cytometry detected in two different types of breast cancer cell conditioned medium(60%)osteogenic cell cycle;and the expression of the Cyclin A2 was detected by Western Blot.2.The expression of protein and m RNA on Notch signaling pathway in osteoblast of Notch1,Jagged1,Hes1 in two different types of breast cancer cell(MDA-MB-231,MCF-7)conditioned medium(0,20%,40%,60%,80%,100%)were detected by Western Blot and q RT-PCR.At the same time,and two different types of breast cancer cell conditioned medium(60%)by DAPT(Notch inhibitor)expression of protein and m RNA processing Notch signal pathway in osteoblast of Notch1,Jagged1,Hes1 were detected.Results:1? MDA-MB-231,MCF-7 in breast cancer cell conditioned medium at different concentrations(0,20%,40%,60%,80%,100%)can inhibit the proliferation of osteoblasts;Two types of MDA-MB-231,MCF-7 breast cancer cell CM with different concentrations(0,20%,40%,60%,80%,100%)treated the osteoblasts,with the increase of concentration and time,the inhibition effect is more obvious,which is positively in the dose effect and time correlation;at the same time,the results of flow cytometry also indicated that both can inhibit the proliferation of osteoblasts,and the main impact of the S phase,RNA synthesis phase;Western Blot results also indicated that both can inhibit the expression of Cyclin A2,and MCF-10 breast cancer cells does not affect the proliferation of osteoblast.To some extent,the above results showed that the appropriate concentration of MDA-MB-231 and MCF-7 breast cancer cell conditioned medium could inhibit the proliferation of osteoblasts.2?MDA-MB-231,MCF-7 in breast cancer cell conditioned medium at different concentrations(0,20%,40%,60%,80%,100%)could up-regulate the expression of protein and m RNA on Notch1,Jagged1 and Hes1 of Notch signal pathway;Two types of MDA-MB-231,MCF-7 breast cancer cell CM with different concentrations(0,20%,40%,60%,80%,100%)treated the osteoblasts,among them,at the concentration of 40% and 60%,the expression of m RNA and protein were the highest,and with the increase of concentration(80%,100%),the osteoblast apoptosis,the expression of protein and m RNA on Notch1,Jagged1 and Hes1 gradually decreased;when adding DAPT(Notch inhibitor),the expression of protein and m RNA of the three were reduced;at the same time.In MDA-MB-231 breast cancer cell in osteoblasts,the expression of protein and m RNA of Hes1 in concentration of 20% began to up-regulate,and at MCF-7 breast cancer cells treated osteoblasts,the expression of protein and m RNA of Hes1 in concentration of 40%;The results showed that the Notch signaling pathway in MDA-MB-231 breast cancer bone metastases is greater than the influence of MCF-7 breast cancer.Conclusion:1.Breast cancer cells(MDA-MB-231,MCF-7)conditioned medium can inhibit proliferation of osteoblast;2.Breast cancer cells(MDA-MB-231,MCF-7)CM could inhibit the proliferation of osteoblasts,which were related with Notch signaling pathways,and there was a more pronounced effect on the associated target gene Hes1 in the Notch signal pathway of MDA-MB-231 CM on osteoblasts.