抑癌基因Kmt2c杂合缺失对小鼠造血系统的影响  

Effect of Tumor Suppressor Gene Kmt2 c Heterozygous Deletion on Hematopoietic System in Mice

作　　者：王雪 华东宁 周瑾 张岩 邢彩虹[1] WANG Xue;HUA Dong-Ning;ZHOU Jin;ZHANG Yan;XING Cai-Hong(Key Laboratory of Chemical Safety and Health,National Institute for Occupational Health and Poison Control,Chinese Center for Disease Control and Prevention,Beijing 100050,China)

机构地区：[1]中国疾病预防控制中心职业卫生与中毒控制所化学安全与健康重点实验室,北京100050

出　　处：《中国实验血液学杂志》2024年第5期1571-1577,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金项目(82070116)。

摘　　要：目的:探究组蛋白甲基转移酶Kmt2c基因杂合缺失对小鼠血液系统的影响。方法:使用CRISPR/Cas9技术构建Kmt2c基因杂合缺失的模型小鼠,血常规连续监测小鼠全血细胞计数的变化;通过体外集落形成实验探究骨髓细胞的克隆性扩增能力;流式细胞术分析突变小鼠体内原始造血细胞群(长期造血干细胞、短期造血干细胞、多能祖细胞)的比例变化。结果:成功构建Kmt2c基因杂合缺失小鼠(Kmt2c^(+/-))模型,其Kmt2c mRNA表达水平是C57BL/6J小鼠的28%。Kmt2c^(+/-)小鼠骨髓细胞体外集落形成能力随着传代次数的增加而增强,并在第四代时集落数显著高于对照组(P<0.05)。Kmt2c^(+/-)小鼠原始造血细胞群中的长期造血干细胞和短期造血干细胞比例分别为19.6%±3.3%及28.9%±4.9%较对照组的16.9%±2.6%及18.9%±2.5%有增高趋势,但差异无统计学意义(P>0.05)。Kmt2c^(+/-)小鼠的白细胞计数在监测的第12周后逐渐上升在第14周时为(9.8±1.0)×10^(9)/L,显著高于对照组的(7.3±1.4)×10^(9)/L(P<0.05)。结论:Kmt2c^(+/-)小鼠的骨髓细胞具有克隆性扩增的潜能。Objective:To explore the effect of heterozygous deletion of histone methyltransferase Kmt2c gene on the hematological system of mice.Methods:CRISPR/Cas9 technology was used to construct mice model of Kmt2c heterozygous deletion(Kmt2c^(+/-))and the changes of whole blood cell count in mice were continuously monitored by blood routine test.The clonal expansion ability of bone marrow cells was explored by colony formation assay in vitro and the proportion of primitive hematopoietic cells,including long-term hematopoietic stem cell(LT-HSC),short-term hematopoietic stem cell(ST-HSC),and multipotent progenitor cell in mutant mice was analyzed by flow cytometry.Results:Kmt2c^(+/-)mice model was successfully constructed,and the mRNA expression level of Kmt2c was 28%of that of C57BL/6J mice.The colony formation ability of bone marrow cells of Kmt2c^(+/-)mice in vitro increased with the passage times,and the colony number in the fourth generation was significantly higher than that of control group(P<0.05).The proportions of LT-HSC and ST-HSC in the primitive hematopoietic cell population of Kmt2c^(+/-)mice was 19.6%±3.3%and 28.9%±4.9%,respectively,which showed an increasing trend compared with 16.9%±2.6%and 18.9%+2.5%in control group,but the difference was not statistically significant(P>0.05).The white blood cell count of Kmt2c^(+/-)mice gradually increased after 12 weeks of monitoring and reached(9.8±1.0)×10^(9)/L at the 14h week,which was significantly higher than(7.3±1.4)×10^(9)/L of control group(P<0.05).Conclusion:The bone marrow cells of Kmt2c^(+/-)mice have potential of clonal expansion.

关 键 词：Kmt2c基因 骨髓细胞 克隆扩增 流式细胞术 

分 类 号：R551.3[医药卫生—血液循环系统疾病]