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应用平方波脉冲电转法将编码人ＩＬ－６受体的ｃＤＮＡ转入不表达ＩＬ－６Ｒ的ＢＮＭＬ大鼠急性早幼粒细胞白血病ＬＴ１２细胞中，转染细胞经含６００μｇ／ｍｌ的Ｇ４１８半固体。液体二步法培养，筛出Ｇ４１８抗性细胞；ＦＡＣＳ检测显示该细胞与ＦＩＴＣ标记的抗ＩＬ－６Ｒ单抗呈显著的荧光强度；￣（１２５）Ｉ－ＩＬ－６受体结合实验表明，ｋｄ＝４．７ｎｍ，每个细胞ＩＬ－６Ｒ数为４０００左右；Ｎｏｒｔｈｅｒｎ－Ｂｌｏｔ发现该细胞的ＲＮＡ与￣（３２）Ｐ标记的１．５ｋｂ的ＩＬ－６ＲｃＤＮＡ探针呈明显的杂交信号，证明我们已成功地建立了高表达人ＩＬ－６Ｒ的ＬＴ１２－ＩＬ－６Ｒ￣＋的大鼠急性白血病细胞模型。
The human IL-6 receptor-encoding cDNA was transfected into BNML rat acute promyelocytic leukemia LT12 cells that did not express IL-6R by square wave pulse electrotransfer, and transfected cells were transfected with G418 semisolid containing 600 μg/ml. Two-step liquid culture, G418-resistant cells were screened; FACS showed that the cells and FITC-labeled anti-IL-6R monoclonal antibody showed significant fluorescence intensity; ̄(125) I-IL-6 receptor binding experiments showed that kd At 4.7nm, the number of IL-6R per cell was around 4000; Northern-Blot found that the RNA of this cell showed a significant hybridization signal with the (32)P-labeled 1.5kb IL-6R cDNA probe, proving that we had The rat acute leukemia cell model of LT12-IL-6R ̄+ highly expressing human IL-6R was successfully established.