OBJECTIVE To observe the effects expression and activation on biological cancer stem cells. of blocking CD133 gene characteristic of the colon METHODS CD133＋ colon cancer stem cells （CCSCs） were separated from EpCAMhigh CD44＋ CCSCs through fluorescenceactivated cell sorting （FACS）. The proliferation, the capability of spherical cell formation, neoplasia, and the expression of ABCG2 mRNA of CD133＋ CCSCs were observed after the CD133＋ CCSCs were infected with LV-CD133shRNA. CD133 negative cells were isolated from EpCAMhigh CD44＋ CCSCs with FACS, and the CD133 proteins in CD133- cells were detected with Western blot. RESULTS CD133＋ CCSCs were isolated from EpCAMhigh CD44＋ CCSCs using FACS, and they accounted for 89.2% in the stem cells. In the experimental group, after the CD133＋ CCSCs were knocked down by LV-CD133shRNA RNAi, the growth pattern of the cells in the stem cell culture changed into adherent growth from suspended growth, and couldn＇t generate spherical cells. Results of MTT assay showed that the CD133＋ CCSCs infected with LV-CD133shRNA grew slowly, compared to the cells in the control groups. There was a decrease in the cloning efficiency. The infected cells were transplanted into the BALB/c nude mice. During the observation, no neoplasia was found in the CD133＋ cells infected with LV-CD133shRNA. The level of ABCG2 mRNA expression was lowered greatly （P 〈 0.01）. CD133- cells were obtained from the EpCAMhigh CD44＋ CCSCs using FACS, in which the expression of CD133 protein was positive. CONCLUSION CD133 retains the biological characteristics of the colon cancer stem cells.
目的:探讨结直肠癌耐药细胞增殖及侵袭力与Na^+,K^+-ATP酶活性及其β1亚单位和P糖蛋白(P-gp)表达的关系,及哇巴因增加化疗敏感性的可能机制。方法:以人结直肠癌亲本细胞(SW480)和耐奥沙利铂细胞(SW480/OxR)为研究对象,采用MTS法、Transwell小室、生化酶学、实时定量PCR(Real time quantitative,RT-PCR)及流式细胞技术等方法比较SW480细胞与SW480/OxR细胞的增殖及侵袭力、Na^+,K^+-ATP酶活性及其β_1亚单位和P-gp表达的差异,观察哇巴因对SW480/OxR细胞上述指标的影响。结果:与SW480细胞比较,SW480/OxR细胞增殖活力无明显变化(P>0.05),而细胞侵袭力却显著增加,Na^+,K^+-ATP酶活性下降,β_1亚单位表达下调,P-gp表达上调(P均<0.01);哇巴因能显著抑制SW480/OxR细胞增殖活力,减弱其侵袭力,下调SW480/OxR细胞P-gp蛋白表达,上调SW480/OxR细胞β_1亚单位蛋白表达,增加SW480/OxR细胞Na^+,K^+-ATP酶活性(P均<0.01)。结论:Na^+,K^+-ATP酶活性下降可能源于β_1亚单位表达下调,并参与结直肠癌的耐药;哇巴因能部分逆转结直肠癌耐药细胞MDR,可能与增加Na^+,K^+-ATP酶活性及下调P-gp表达有关。
目的研究分析应用表柔比星联合紫杉醇的新辅助化疗方法治疗三阴性乳腺癌的效果。方法经筛选将昆明医科大学第三附属医院40例三阴性乳腺癌患者做为研究对象,采用应用表柔比星联合紫杉醇的新辅助化疗法进行治疗,分别测定其治疗前后叉头框蛋白A1(fork box protein A1,FOXA1)、乳腺癌易感基因(breast cancer susceptibility gene 1,BRCA1)蛋白表达水平、肿瘤直径大小。结果治疗后,患者的FOXA1、BRCA1蛋白表达水平降低,与治疗前相比差异具有统计学意义(P<0.05);治疗后,患者肿瘤直径减小,与治疗前相比差异具有统计学意义(P<0.01)。结论采用应用表柔比星联合紫杉醇的新辅助化疗,对三阴性乳腺癌患者进行治疗,治疗后疗效显著,FOXA1、BRCA1水平改善,肿瘤直径减小。
