Effect Of Compatibility Of Radix Aconiti Praeparata And Bulbus Fritillaria On Human Breast Cancer

Objective:The present study was designed to investigate the effect of Radix Aconiti Praeparata in combination with Bulbus Fritillaria on breast cancer cells under different drug dosage and combination ratio are observed in vitro.Methods:1.Human breast cancer MDA-MB-231 cells were seeded in culture plates.Set up a system of different concentrations of Radix Aconiti Praeparata(RAP)and BulbusFritillaria(BF)single-use group,combined-use group,as well as compatibility by 1:1,2:1 proportion and negative control group.Effects of BF and RAP on viability of MDA-MB-231 were determined by trypan blue staining method.AQP1 expression in cells were observed after beening treated with BF and RAP through enzyme-linked immunosorbent assay(ELISA).2.The status of the breast cancer cell migration was studied through Wound-healing assay and Transwell assay.The the localization and expression of E-cadherin and actin microfilament were observed under a confocal microscope.3.Set up a system of different concentrations of aconitine and tubeimoside I single-use group,combined-use group,as well as compatibility by '1:1,2:1,1:2 proportion and negative control group for SKBR3 and MDA-MB-231 cells.The antiproliferative activity of tubeimoside I and aconitine was determined through MTS method.The status of migration was also studied through Transwell assay.Results:1.Treatment with BF and/or RAP,no significant difference of viability was observed(P>0.05).However,treatment with BF and concomitant treatment BF and RAP,MDA-MB-231 cells were tumid,and abundant vacuolations in cytoplasm were detected by optics microscope,and the expression of aquaporin-1 was decreased(P<0.01).There was no significently influnce of RAP on the viability,cell damage and aquaporin-1 expression of MDA-MB-231 cells.2.(1)Wound-healing assay:The group of BF(62.5?g/mL)and group of concomitant treatment BF and RAP(62.5:31.3?g/mL)down regulated SKBR3 cell migration,while RAP(31.3?g/mL)up regulated cell migration,as compared to the control cells.(2)Transwell assay:The number of RAP-treated cells increased significantly compared to the control cells(P<0.05).The number of BF-treated cells and concomitant treatment BF and RAP decreased significantly compared to the control cells(P<0.01)and RAP-treated cells(P<0.01).There were no difference of cell migration between concomitant treatment and BF treatment in MDA-MB-231 cell(P>0.05).However,concomitant treatment BF and RAP exert synergic effects to inhibit the migration of SKBR3 cells(P<0.01).(3)Confocal microscopy:RAP-treated cells showed pseudopodia formation and and actin filaments were partially accumulated at the leading edge of ruffled cell membranes forming lamellipodia.However,actin filaments in BF treated cells were accumulated distributed in the cytoplasm.Combined treatment with BF and RAP,SKBR3 cells showed abnormal feature with the distribution of actin filaments.3.The number of tubeimoside I-treated cells was significantly less than the control cells and aconitine-treated cells(P<0.05)when the tubeimoside I dose was 10?g/mL.Tubeimoside I acted synergistically with aconitine in inhibiting the proliferation of SKBR3 cells in compatibility of 10:10?10:20?g/mL(P<0.05).Tubeimoside I acted synergistically with aconitine in inhibiting MDA-MB-231 cell migration in compatibility of 5:2.5?g/mL,while in inhibiting SKBR3 cell migration in compatibility of 3:1.5 and 5:2.5?g/mL.There are significant differences between concomitant treatment tubeimoside I and aconitine and two drugs alone(P<0.05).Conclusions:1.With a appropriate compatible ratio,BF and RAP did not increase the cytotoxic effect of breast cancer cells.2.BF down regulated SKBR3 cell migration,while RAP up regulated MDA-MB-231 cell migration.BF acted synergistically with RAP in inhibiting the proliferation of SKBR3 cells.3.Tubeimoside I acted synergistically with aconitine in inhibiting the proliferation and migration of breast cancer cells.4.Wann and resolve cold-phlegm is a vital therapeutic principle for breast cancer.