| 【Objectives】:In this study, using immunomagnetic beads sorting technique selected from breast cancer cell line MCF-7carve the breast cancer stem cell subsets (CD44+CD24"), And planted subcutaneously into nude mice axillary. After randomization, To pour the fluid that the corresponding single Traditional Chinese medicine (Ginseng, Astragalus, Angelica, Ganoderma, Aspartate) decoction into stomach respectively. To observe the effect of righting anticancer Traditional Chinese medicine on tumorigenicity of human breast cancer stem cell CD44+CD24-subsets, and explore its mechanism by detect of immune system-related factors and tumor associated gene expression.【Methods】:1. Separate the breast cancer stem cell subsets (CD44+CD24-) from breast cancer cell MCF-7with magnetic cell sorting technology.2. Planted the breast cancer stem cell subsets (CD44+CD24-) subcutaneously into nude mice armpit, these nude mice were divided into seven groups based on the principle of randomization:the ginseng group, the astragalus group, the angelica group, the ganoderma group, the radix asparagi group, the epirubicin group, the model group.3. These nude mice of various traditional Chinese medicine group were poured the fluid that the corresponding single Traditional Chinese medicine (Ginseng, Astragalus, Angelica, Ganoderma, Aspartate) decoction into stomach respectively, once a day; The model group were poured the same amount of saline; epirubicin group, intraperitoneal injection, once three weeks,12weeks of continuous administration.4. Observe tumor formation time and record the tumor volume. After administration, collecting blood and mice were sacrificed, peel the tumor tissue, measure tumor weight and calculate the inhibitory rate.5. Detected TNF-a, IL-2concentration in the serum of nude mouse in each experimental group through Elisa method.6. Detected the relative expression levels of CD24mRNA、CD44mRNA in tumor tissues of nude mouse in each experimental group through RT-PCR.[Results]:1. The tumor formation time of nude mouse in each group:The various traditional Chinese medicine group was extended significantly than the epirubicin group and model group, the difference was statistically significant (P <0.05).2. The average tumor volume and the average tumor weight of nude mouse in each group:The various traditional Chinese medicine group and the epirubicin group was significantly lower than the model group, the difference was statistically significant (P<0.05); In the six treatment groups, The tumor inhibition rate of the ginseng group is highest, up to68.24%; The tumor inhibition rate is silimar between the remaining four traditional Chinese medicine group and epirubicin group.3. The concentration of TNF-a in the serum of nude mouse in each group: The concentration of TNF-a in various traditional Chinese medicine group and epirubicin group reduced to varying degrees than the model group and epirubicin group, the difference was statistically significant (P<0.05); The ginseng group have a lower concentration than the epirubicin group, the difference was statistically significant (P<0.05); The difference was not statistically significant between the remaining four traditional Chinese medicine group and the epirubicin group (P>0.05).4. The concentration of IL-2in the serum of nude mouse in each group: The various traditional Chinese medicine group and epirubicin group reduced to varying degrees than the model group and epirubicin group, the difference was statistically significant (P<0.05);The ganoderma group have a higher concentration than the epirubicin group, the difference was statistically significant (P<0.05); The difference was not statistically significant between the remaining four traditional Chinese medicine group and the epirubicin group (P>0.05).5. The relative expression levels of CD24mRNA in tumor tissues of nude mouse in each group:The relative expression levels of CD24mRNA in tumor tissues of nude mouse in various traditional Chinese medicine group and epirubicin group reduced to varying degrees than the model group, the difference was statistically significant (P<0.05). The difference was not statistically significant between various traditional Chinese medicine group and epirubicin group (P>0.05).6. The relative expression levels of CD44mRNA in tumor tissues of nude mouse in each group:The relative expression levels of CD44mRNA in tumor tissues of nude mouse in various traditional Chinese medicine group and epirubicin group reduced to varying degrees than the model group, the difference was statistically significant (P<0.05). The difference was not statistically significant between various traditional Chinese medicine group and epirubicin group (P>0.05).【Conclusions】:1. Five righting class of anti-cancer traditional Chinese medicine can effectively extend the time of the tumorigenicity of human breast cancer stem cells, at the same time,can effectively inhibit tumor growth. Show that the centralizer class of anti-cancer traditional Chinese medicine can effectively suppress the tumorigenicity of human breast cancer stem cells.2. The centralizer class of anti-cancer traditional Chinese medicine can effectively reduce the concentration of TNF-a in the serum of nude mouse with breast cancer, increase the concentration of IL-2in the serum of nude mouse with breast cancer, and reduce the relative expression levels of CD24mRNA、 CD44mRNA in transplanted tumors tissues of nude mouse. Show that these may be part of the centralizer class of anti-cancer traditional Chinese medicine inhibition of tumor stem cells into tumor mechanism. 3. This study provide new areas and ideas of research in human breast cancer tumor stem cell tumorigenicity, further enhance the level and depth of research in the study on the centralizer class of anti-cancer traditional Chinese medicine inhibition of tumor. |
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