【摘要】 目的 探讨集落刺激因子1(CSF1)在子宫内膜癌细胞系中的表达,为靶向治疗子宫内膜癌提供新的治疗思路。方法 采用实时荧光定量聚合酶链式反应法、免疫印迹方法检测CSF1在子宫内膜癌Ishikawa细胞系中的表达情况以高表达CSF1的Ishikawa细胞作为研究对象,利用特异性靶向CSF1基因的shRNA慢病毒敲除系统包装病毒,并感染Ishikawa细胞,使用流式细胞分选技术获得CSF1稳定敲除细胞株,并分别在mRNA、蛋白水平检测shRNA敲除效率,同时监测CSF1敲除后不同时间点细胞增殖及凋亡水平。结果 CSF1在子宫内膜癌Ishikawa细胞中高表达利用shRNA慢病毒敲除系统,抑制CSF1在Ishikawa细胞中的表达可通过上调凋亡促进因子BAX,导致细胞增殖明显减缓,并诱导细胞凋亡(均P <0.05)。结论 CSF1在子宫内膜癌细胞系中高表达,抑制CSF1的表达可引起细胞大量凋亡,揭示CSF1可能对子宫内膜癌发生及发展具有正向调控作用。更多还原

【Abstract】 Objective To explore the expression,function and related mechanisms of colony stimulating factor 1(CSF1)in human endometrial cancer cell line and to provide a new strategy for targeted therapy of endometrial cancer.Methods Real-time quantitative PCR and Western blot were used to detect the expression of CSF1 in ISHIKAWA cells.Using Ishikawa cells with high expression of CSF1 as the research object,a shRNA lentivirus knockout system specifically targeting the CSF1 gene was used to package the virus and infect Ishikawa cells,and stable knockdown cell lines of CSF1were obtained by flow cytometry.The knockdown efficiency in the mRNA level and protein level were detected,while cell proliferation and apoptosis levels were monitored at different time points after CSF1 knockout.Results Real-Time PCR and Western blot results showed that CSF1 was highly expressed in ISHIKAWA cells.Using shRNA lentivirus knockdown system could significantly knockdown the expression of CSF1 via upregulating the pro-apoptotic factor BAX,which could inhibit cell proliferation dramatically and induce cell apoptosis(all P <0.05).Conclusions CSF1 can be highly expressed in endometrial cell lines.Suppressed CSF1 expression can significantly induce cell apopto sis,which uncovered that CSF1may have a positive effect on the occurrence and development of endometrial cancer.更多还原