The Antitumor Effect Of Different Combination Ways Of Cyclophosphamide With Zoledronic Acid On MDA-MB-231 Breast Cancer Cells

Breast cancer is the most common malignancy in women,the incidence and the mortality rate of breast cancer has been the first female tumor in the world.Moreover,breast cancer as a high heterogeneity so current chemotherapy,radiotherapy,endocrine therapy and targeted therapy are difficult to achieve ideal therapeutic effect,expecially triple-negative breast cancer(TNBC)which the phenotypes of estrogen receptor(ER),progesterone receptor,(PR)and human epidermal growth factor receptor 2(HER-2)are both negative,there is no effective treatments.Therefore,it is urgent to develop new drugs and invent more effective combination drug programs so that improving the effect of breast cancer treatment.Objective:zoledronic acid,who has the inhibition effect of bone metastasis,has been proved that it has synergistic antitumor role with imatinib mesylate on lukemia.Meanwhile,it also demonstrated that it can prolong the survival time of patients with breast cancer by combination with endocrine drugs.However,the antitumor effect of zoledronic acid combined with chemotherapy drugs on breast cancer has not been reported before.So in this study,we performed the antitumor study of different combination ways of zoledronic acid with cyclophosphamide combined on the human three-negative breast cancer MDA-MB-231 cells,so that evaluating the effect of these combination and offering the best combination order of zoledronic acid with first-line chemotherapy drugs and provide new experimental basis for the drug and clinical drug regimen for the treatment of triple negative breast cells.Methods:the experiments were divided into control groups,monotherapy cyclophosphamide groups,monosulfonyl phosphate groups,simultaneous administration of zoledronic acid and cyclophosphamide groups,cyclophosphamide sequence to zoledronic acid groups,and zoledronic acid sequence to cyclophosphamide groups.Secondly,the antitumor effects of different concentrations of zoledronic acid or cyclophosphamide on MDA-MB-231 cells were observed and measured by microscope and CCK-8 assay so that analyzing the best optional concentration.Meanwhile,the antitumor effects and anti-metastasis effects of the optimal doses of cyclophosphamide and zoledronic acid on MDA-MB-231 cells were respectively evaluated by morphology observation,CCK8 assays,Calcein-AM/PI assays and transwell assys.Results: Firstly,The proliferation of breast cancer MDA-MB-231 cells was inhibited by different concentrations of zoledronic acid and cyclophosphamide in different time compared with that of control(P<0.05),and the inhibition role of zoledronic acid and cyclophosphamide are both depended on doses and time,meanwhile,we found the IC50 of zoledronic acid and cyclophosphamide on MDA-MB-231 cells were respectively 1 ?mol/L and 12.5 ?mol/L in 72 h,so the antitumor effect of cyclophosphamide was better than the zoledronic acid.To better know the combination effect of cyclophosphamide with zoledronic acid on breast cancer MDA-MB-231,we choose 0.1 ?mol/L Cyclophosphamide combined with 3.125 ?mol/L zoledronic acid in different sequence to stimulate the MDA-MB-231 cells,the results of CCK8 assay,Calcein-AM/PI assays and transwell assays were shown the antitumor effects and anti-metastasis role of Cyclophosphamide added with zoledronic later were obvious stronger than that of others.Conclusion: Zoledronic acid has synergistical effects on cyclophosphamide and this effects can be affected by the combination sequence.In those combination,the cyclophosphamide first and then combined with zoledronic acid later showed stronger antitumor ability and anti-metastasis ability,these results suggested that this drugs combination not only can kill tumor cells,but also prevent the development of tumor metastasis,hence this project would improve the theraputic effect of metastic bread cancer.