Research On The Effect And Mechanism Of PA-MSHA On Breast Cancer

Part I A Clinical Research on Pseudomonas Aeruginosa Injection for the Treatment of Malignant Ulcer of Breast CancerObjective:To evaluate the effectiveness and adverse reaction of pseudomonas aeruginosa injection (PA-MSHA) for the treatment of malignant ulcer of breast cancer.Methods:Patients with malignant ulcer of breast cancer were randomly divided into two groups such as the control group receiving chemotherapy alone and the treatment group receving chemotherapy combined with PA-MSHA. Tumor size and ulcer area were measured before and after treatment. The adverse reactions were also evaluated following with the process of the therapy.Results:Contrast with the control group(33.3%), the treatment group(85.7%) could promote malignant ulcer area growing downwards(P<0.05), but there was no statistical significance(P>0.05) about breast cancer tumor between the treatment group(57.1%) and the control group(33.3%).Conclusion:Administration of PA-MSHA is effective in the treatment of malignant ulcer in breast cancer without obvious adverse reactions. PartⅡKilling Mechanism Research on Pseudomonas Aeruginosa MSHA (PA-MSHA) Injection in Breast Cancer in VitroObjective:To investigate the effects of PA-MSHA on the apoptosis and proliferation of human breast cancer cells (MCF-7) cultured in vitro.Methods:With culture in vitro of MCF-7 cells being in logarithmic phase, then divided them into two groups such as the expertiment group and the control group. The experiment group were given PA-MSHA injection, and the contro group were mitte tales doses of RPMI 1640 culture medium. With MTT colorimetric method detecting the inhibition effect of PA-MSHA on MCF-7 cultured in vitro, and flow cytometry dectected the influence of PA-MSHA affecting MCF-7 apoptosis and cell cycle. Then used Hoechst fluorescent staining to observe it induce the morphology chage of MCF-7 cultured in vitro.Results:Contrast with the control group, different densities of PA-MSHA all could inhibit MCF-7 cell proliferation(P<0.05), promote MCF-7 cell apoptosis (PA-MSHA group apotosis rate:2.14±0.246, the control group rate:1.19±0.178, P＜0.01), raised G0/G1 stage cell ratio(PA-MSHA group apotosis ratio:73.00±7.10, the control group ratio:49.83±9.83) of MCF-7 cell and cut down the percentage of G2/M and S stages. Fluorescence microscope detecting showed the obvious apoptosis image of MCF-7 cell.Conclusion:PA-MSHA mighg through inducing MCF-7 apoptosis, played inhibition effect in the G0/G1 stage of cell cycle, thus inhibited MCF-7 cell proliferation. Part III Therapeutic Action and Mechanism Study on PA-MSHATreating Breast cancer in VivoObjective:1. To investigate the effects of PA-MSHA for the treatment of brease cancer in high spontaneous metastasis breast cancer of mouse model, and explore its mechanism of action.2. To study the effects of PA-MSHA on the immune function in patients with advanced stage breast cancer.Methods:Use of 4T1 cell suspension of cell strains building up high spontaneous metastasize breast cancer mice model, divided mice with tumor into four groups, such as negative control group(A group), CTX treatment group(B group), PA-MSHA group(C group) and therapeutic alliance group (D group). Compared tumor size and weight, life span, pulmonary metastasis rate among groups after treatments. And use of flow cytometry detection method compared peripheral blood lymphocyte subpopulation among different groups, applied with cytometric bead array(CBA) detecting TNF-a, IFN-y, IL-5, IL-4, IL-2 content of peripheral blood cytokines in mices. Pearsonian correlation analysis was used to investigate the relationship between peripheral blood lymphocyte subpopulation, TNF-a, IFN-y, IL-5, IL-4, IL-2 and survival time.Of 14 patients with advanced stage breast cancer were randomly divided into two groups as chemotherapy group and chemotherapy associated with PA-MSHA group. Both groups were treated with chemotherapy and the PA-MSHA group were injected with PA-MSHA by oneself, while the two groups were treated with every other day. The contents of T cells(CD3+, CD4+, CD8+) phenotype in blood was detected by flow cytometry (FCM), and computed the ratio of CD4+/CD8+.Results:Compared with the negative group, the treatment groups had smaller size of tumor and lighter weight of tumor, but there were no statistical significance between the CTX group and PA-MSHA group (P=0.160), both larger than therapeutic alliance group (P<0.05). Compared with Pulmonary metastasis rate, the negative group was much higher than the therapeutic alliance group (P<0.05), but there were no statistical significance among other groups (P>0.05). Contrast with life span, there was no significance between the PA-MSHA group and the CTX group (P=0.056), both higher than the negative group (P<0.05),both lower than the therapeutic alliance group(P<0.05). Compared with spleen index, the control group was smaller than the other four groups (P<0.05), the therapeutic alliance group was higher than the other four groups(P<0.05), but there were no statistical significance between the CTX group and the PA-MSHA group (P=0.352). Contrast with the negative group, PA-MSHA could elevate the ratio of CD4 cells of peripheral blood T lymphocytes, cut down the ratio of CD8 cells of peripheral blood T lymphocytes, and raise the ratio of CD4/CD8 (P<0.05). The level of IL2 was higher in PA-MSHA group (P<0.05), and the levels of TNF-a and IFN-y were higher in PA-MSHA& CTX group (P< 0.05). There was good positive correlation between CD4+and survival time (r=0.714, P＜0.01), but there was negative correlation between CD8+and survival time r=0.790, P＜0.01). There was good positive correlation between CD4+/CD8+ and survival time (r=0.781, P<0.01), and there was positive correlation between IL-2 and survival time (r=0.552,P<0.05). Other indexes didn't exist correlation with survival time(P>0.05).In the clinical research, there was no statistical significance between the PA-MSHA group and the control group(P>0.05). The CD4+T cells and CD4+/CD8+ ratios of the experiment group were higher than the control group(P<0.05), but the CD8+T cells ratio of the experitment group was lower than the control group (P< 0.05).Conclusions:1.There are immune imbalance in the high spontaneous metastasis breast cancer mouse model that set up with 4T1 cell.2. The PA-MSHA could change the immune state in the tumor-bearing mices so as to treat breast cancer, and prolong the lifetime. And it was more effective together with CTX.3. PA-MSHA could enhance cellular immune function in advanced stage of breast cancer patients.