Zhu Yufeng, Wang Yuanzhan, Meng Fanyi
Laboratory Animal Research Center of Nanfang Hospital, the Southern Medical University, Guangzhou 510515, China.
Zhonghua Xue Ye Xue Za Zhi. 2015 Mar;36(3):221-4. doi: 10.3760/cma.j.issn.0253-2727.2015.03.010.
To construct the P210(T315I-BCR/ABL) transgenic mice model.
The transgenic vector in which the P210(T315I-BCR/ABL) gene and eGFP gene was derived by APN/CD13 promoter was constructed and microinjected into the single-cell fertilized eggs of C57 mice. Transgene integration was conformed by PCR genotyping and P210(T315I-BCR/ABL) expression levels was evaluated by RT-PCR. The CML phenotype was confirmed by blood routine examination, Wright's staining for peripheral blood and bone marrow smears, HE staining for organs of transgenic mice.
Three transgenic mice lines with high expression of P210(T315I-BCR/ABL) gene and eGFP gene was selected. Compared with the wild type mice, the levels of WBC, platelet and neutrophil granulocyte of transgenic mice began to increase gradually at 2 months, and increase to 23.9×10⁹/L, 4 136×10⁹/L, and 74.6% respectively at 6 months. The remarkable hyperplasia of granulocytes was seen in the peripheral blood and bone marrow smears with splenomegaly infiltrated by leukemic cells.
The P210(T315I-BCR/ABL) transgenic mice was constructed and provided a model to explore the mechanism of T315I CML and screen out the drug for T315 CML patient.
构建P210(T315I-BCR/ABL)转基因小鼠模型。
构建由APN/CD13启动子驱动P210(T315I-BCR/ABL)基因和eGFP基因的转基因载体,并显微注射到C57小鼠的单细胞受精卵中。通过PCR基因分型确认转基因整合情况,用RT-PCR评估P210(T315I-BCR/ABL)表达水平。通过血常规检查、外周血和骨髓涂片的瑞氏染色、转基因小鼠器官的HE染色来确认慢性髓性白血病(CML)表型。
筛选出3个P210(T315I-BCR/ABL)基因和eGFP基因高表达的转基因小鼠品系。与野生型小鼠相比,转基因小鼠的白细胞、血小板和中性粒细胞水平在2个月时开始逐渐升高,6个月时分别升至23.9×10⁹/L、4 136×10⁹/L和74.6%。在外周血和骨髓涂片中可见粒细胞显著增生,伴有脾肿大且有白血病细胞浸润。
成功构建了P210(T315I-BCR/ABL)转基因小鼠,为探索T315I慢性髓性白血病的发病机制及筛选针对T315慢性髓性白血病患者的药物提供了模型。