| Part 1.Clinical efficacy evaluation of liver regulating,kidney tonifying and accumulation eliminating therapy for patients with advanced breast cancer in stable phaseObjective:To explore the effect of TCM liver regulating,kidney tonifying and accumulation eliminating therapy on disease progression,TCM symptoms,quality of life and immune function of patients with advanced breast cancer in stable phase.Methods:A prospective cohort design was used to collect 100 patients with advanced breast cancer whose condition was stable after first-line chemotherapy in our hospital from September 2014 to September 2017.They were divided into control group and observation group according to whether they received the treatment of liver regulating,kidney tonifying and accumulation eliminating therapy of traditional Chinese medicine.The control group was treated with routine western medicine,while the observation group was treated with traditional Chinese medicine for liver regulating and kidney tonifying and accumulation eliminating on the basis of western medicine.After treatment,the progression free survival(PFS),TCM symptom score,Kanofsky score(KPS),Hamilton Depression Scale(HAMD),peripheral blood cell immunity and humoral immunity levels of the two groups were analyzed and compared.Results:A total of 95 effective cases were collected,47 in the control group and 48 in the observation group.(1)Compared with PFS of two groups of patients,M-PFS of control group is 5.90months,that of observation group is 8.00 months,and that of observation group is 2.1months longer than that of control group(P<0.05);There was no significant difference in PFS between the two groups of ER(-)/PR(-)patients(P>0.05);Compared with PFS of ER(+)/PR(+)patients in the two groups,M-PFS in the control group is 6.00 months,that in the observation group is 10.40 months,and that in the observation group is 4.4months longer than that in the control group(P<0.05);Compared with the PFS of Her-2(-)patients in the two groups,the M-PFS in the control group is 6.20 months,that in the observation group is 10.60 months,and that in the observation group is 4.4 months longer than that in the control group(P<0.01);There was no significant difference in PFS between the two groups of Her-2(+)patients(P>0.05).(2)Compared with traditional Chinese medicine symptom scores of the two groups of patients,the observation group’s traditional Chinese medicine emotional depression,insomnia,vexation and irritability,spontaneous perspiration and night sweat,five vexation and fever,soreness of waist and knees,xerostomia symptom scores and total traditional Chinese medicine symptom scores after treatment are significantly lower than those before treatment,and are significantly lower than those of the control group after treatment(P<0.05,P<0.01).(3)Compared with KPS scores of two groups of patients,the improvement rate of the observation group was 45.8%,and that of the control group was 21.3%,the observation group was significantly higher than the control group(P<0.01).The deterioration rate of the observation group was 6.3%,and that of the control group was19.1%,the observation group was significantly lower than the control group(P<0.01).(4)Compared with HAMD scores of the two groups,the effective rate of the observation group was 77.1%,and that of the control group was 57.4%,which was significantly higher than that of the control group(P<0.05).(5)Compared with the cellular immunity and humoral immunity indexes of the two groups of patients,the CD3~+level,CD4~+level,CD4~+/CD8~+ratio,NK cell level and immunoglobulin Ig G,Ig M and Ig A contents of the observation group after treatment are significantly higher than those before treatment,and are significantly higher than those of the control group after treatment(P<0.05,P<0.01).Conclusions:TCM therapy of liver-regulating,kidney-tonifying and accumulation-eliminating therapy can delay the progress of disease,improve the TCM symptoms,quality of life and immune function of patients with advanced stable breast cancer in stable stage.Part 2.Effect of Curdione on Migration and Invasion of Breast Cancer Cells and Its MechanismObjective:To investigate the effect of curdione on the migration and invasion of human breast cancer MDA-MB-231 and MCF-7 cells and its possible mechanism.Methods: CCK-8 assay tests the effect of curdione on the growth activity of MDAMB-231 and MCF-7 cells,and to determine the concentration of drugs in subsequent experiments;Cell adhesion assay was used to detect the effect of curdione on the adhesion ability of MDA-MB-231 and MCF-7 cells;Wound healing assay was used to detect the effect of curdione on the scratch migration ability of MDA-MB-231 and MCF-7 cells;Transwell assay was used to detect the effect of curdione on the migration and invasion of MDA-MB-231 and MCF-7 cells;Western blot was used to detect the effect of curdione on the expression of key proteins ERK,p38,Akt and phosphorylation levels in MAPK and Akt signaling pathways in MDA-MB-231 and MCF-7 cells;Western blot and Real-time PCR were used to detect the effect of curdione on the expression of MMP-2,MMP-9 protein and m RNA in MDA-MB-231 and MCF-7 cells.Results:(1)CCK-8 results showed that when the concentration of curdione≤50μmol/L had no obvious effect on the growth activity of MDA-MB-231 and MCF-7 cells,when the concentration of curdione≥100μmol/L had obvious inhibitory effect on the growth activity of MDA-MB-231 and MCF-7 cells(P<0.05,P<0.01),which was timedependent and concentration-dependent.Curdione concentrations(12.5μmol/L,25μmol/L,50μmol/L)that had no significant effect on cell growth activity were selected for subsequent experiments.(2)The results of cell adhesion experiments showed that the cell adhesion rate of MDA-MB-231 and MCF-7 cells was significantly lower than that of the blank control group(P < 0.01)after 24 hours of intervention with 12.5μmol/L,25μmol/L and 50μmol/L concentration curdione,and was concentration dependent within a certain range.(3)Wound healing assay results show that MDA-MB-231 and MCF-7 cells have significantly lower scratch migration rate than the blank control group(P<0.05,P<0.01)after continuous intervention of 12.5μmol/L,25μmol/L and 50μmol/L concentration curdione for 24 h and 48 h,and show concentration dependence within a certain range.(4)Transwell assay results showed that after 24 hours of intervention with 12.5μmol/L,25μmol/L and 50 μmol/L concentration curdione,the number of migrated and invaded cells was significantly lower than that of the blank control group(P<0.05,P<0.01),and was concentration dependent within a certain range.(5)Western blot results showed that,compared with the blank control group,curdione with concentrations of 12.5μmol /L,25μmol /L,and 50μmol /L could significantly inhibit the phosphorylation levels of ERK,p38,and Akt proteins in MDA-MB-231 and MCF-7 cells(P<0.05,P<0.01)in a concentration-dependent manner within a certain range.(6)The results of Western blot and Real-time PCR showed that,compared with the blank control group,curdione with concentrations of 12.5μmol/L,25μmol/L and 50μmol/L could significantly reduce the expression levels of MMP-2,MMP-9 protein and m RNA in MDA-MB-231 and MCF-7 cells(P<0.01),and showed concentration dependence within a certain range.Conclusions: Curdione can significantly reduce the migration and invasion ability of MDA-MB-231 and MCF-7 in human breast cancer.The mechanism may be related to down-regulating the phosphorylation levels of key proteins ERK,p38 and Akt in MAPK and Akt signaling pathways,and further down-regulating the expression levels of MMP-2 and MMP-9. |
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