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目的分析大肠癌始动细胞群锌指蛋白表达谱变化信息。方法采用无血清悬浮培养结合CD133抗体标记磁珠分选获得Colo205来源大肠癌始动细胞,高通量测序分析180个锌指蛋白基因mRNA表达谱变化,并筛选差异表达基因,利用荧光定量PCR技术对部分差异表达基因进行表达变化验证。结果与CD133-细胞群相比,CD133+细胞群有56个锌指蛋白基因表达下调,42个锌指蛋白表达上调;与Colo205细胞群相比,CD133+细胞群有58个锌指蛋白基因表达下调,40个锌指蛋白表达上调;与Colo205细胞群相比,CD133-细胞群有79个锌指蛋白基因表达下调,46个锌指蛋白表达上调,定量PCR检测结果与表达谱测序数据表现出趋势一致性。CD133-细胞群与Colo205细胞群相比,在CD133+细胞群中有29个基因的表达均表现出了较大幅度的改变,其中有11个基因在前两种细胞群中表现为表达上调,13个基因表现为下调。结论无血清培养基培养结合CD133磁珠分选Colo205细胞获得的CD133+细胞亚群中众多与肿瘤发生、发展有关的锌指蛋白基因在CD133+细胞群中的表达发生改变。
Objective To analyze the changes of zinc finger protein expression profile of primary colorectal cancer cell population. Methods Colo205 primary colorectal cancer cells were obtained by serum-free suspension culture combined with labeled beads of CD133 antibody. The mRNA expression profiles of 180 zinc finger proteins were analyzed by high-throughput sequencing, and the differentially expressed genes were screened by fluorescence quantitative PCR The expression of some differentially expressed genes was verified. Results Compared with the CD133 cell population, 56 zinc finger proteins were down-regulated and 42 zinc finger proteins were up-regulated in the CD133 + cell population. Compared with the Colo205 cell population, 58 zinc finger proteins were down-regulated in the CD133 + 40 zinc finger proteins were upregulated. Compared with Colo205 cell population, 79 zinc finger protein genes were down-regulated and 46 zinc finger proteins were up-regulated in CD133- cell population. The results of quantitative PCR showed consistent trend with those of expression profiling data Sex. The expression of 29 genes in the CD133 + cell population showed a significant change compared with the Colo205 cell population, of which 11 genes were up-regulated in the first two cell populations 13 A gene showed a decrease. Conclusion Many of the CD133 + cell subpopulations isolated from CD153 cells cultured in serum-free medium with CD133 magnetic beads change the expression of many zinc finger proteins involved in tumorigenesis and development in the CD133 + cell population.