【摘要】 为了探究敲除长非编码RNA SNHG16对人慢性髓系白血病K562细胞的影响,我们利用CRISPR/Cas9技术在K562细胞中敲除SNHG16基因,通过流式分选获得单细胞,经扩增培养、基因组PCR鉴定、测序鉴定后,获得SNHG16杂合和纯合敲除株;通过Wright-Giemsa染色、MTS检测、流式分析和qRT-PCR分别检测了SNHG16敲除后对K562细胞形态、增殖、细胞表面标志蛋白及红系分化调控因子的影响。实验结果显示,敲除SNHG16后不影响K562细胞的形态和增殖,显著促进了K562细胞表面标志蛋白CD235a和红系分化调控因子的表达水平。该研究表明,长非编码RNA SNHG16不影响K562细胞的增殖,但SNHG16对K562细胞表面标志蛋白CD235a的表达水平有一定的调控作用。更多还原

【Abstract】 The aim of this work was to investigate the effect of long noncoding RNA SNHG16 knock-out on human chronic myeloid leukemia cells（K562 cells）. The SNHG16 gene in K562 cells was knocked out by using CRISPR/Cas9 system. Single cells were gained by flow cytometric sorting technique. SNHG16+/– and SNHG16^–/– cell lines were gained after culture, identification of PCR and sequencing. The effects of SNHG16 depletion on morphology, proliferation, cell surface marker and erythroid transcription factors of K562 cells were detected by Wright-Giemsa staining, MTS assay, flow cytometry, and qRT-PCR. The results indicated that SNHG16 depletion did not affect the morphology and proliferation of K562 cells, but promoted the expression level of K562 cell surface marker CD235 a and erythroid transcription factors. In conclusion, long noncoding RNA SNHG16 does not affect the proliferation of K562 cells, but SNHG16 plays a role in the regulation of K562 cell surface marker protein CD235 a.更多还原