FKBP-RAP-FRB系统转运BCR-ABL入核对K562细胞的增殖抑制效应

BCR-ABL融合蛋白是慢性粒细胞白血病（chronic myeloid leukemia，CML）发病的基础。其中，BCR-ABL只能定位于细胞浆、不能易位至细胞核是其致病的关键因素。因此，转运BCR—ABL入核可能是治疗CML的潜在方法。该研究利用基因重组技术，构建HA-2FKBP-ABD（HF2A）和FLAG-3NLS—FRB＊（FN3R）重组腺病毒，与雷帕霉素类似物（Rapamycin analog）同组成FKBP-RAP-FRB系统，转运K562细胞胞浆中的BCR—ABL癌蛋白至细胞核，并探究其对K562细胞增殖的影响。结果显示，成功构建了高滴度的重组腺病毒，Westernblot证实目的蛋白在K562细胞内成功表达。FKBP—RAP—FRB系统可通过转运BCR—ABLA入核。抑制K562细胞生长和克隆形成的能力。结果揭示，FKBP-RAP—FRB系统转运BCR—ABL入核有望为CML提供新的治疗手段。

Inhibitory Effect on Proliferation of K562 Cells by Transporting BCR-ABL into Nucleus with FKBP-RAP-FRB System

BCR-ABL fusion protein is demonstrated as the pathogenesis of chronic myeloid leukemia （CML）. Failure of cytoplasmic BCR-ABL translocating into nucleus plays a key role. Therefore, transporting BCR- ABL into nucleus may be a potential therapeutic approach of CML. In this study, HA-2FKBP-ABD （HF2A） and FLAG-3NLS-FRB＊ （FN3R） recombinant adenovirus were constructed by recombinant DNA technology. HF2A, FN3R, and rapamycin analog constituted FKBP-RAP-FRB system, which was used to transport cytoplasmic BCR- ABL oncoprotein into nucleus. By this mean, the effect on the proliferation of K562 cells was detected. Our results showed that high titer of recombinant adenovirus were successfully constructed, and the target protein was success- fully expressed in K562 cells confirmed by Western blot. FKBP-RAP-FRB system could inhibit growth and colony formation of K562 cells by transporting BCR-ABL into nucleus. These results revealed that FKBP-RAP-FRB sys- tem transporting BCR-ABL into nucleus could provide new insights for the treatment of CML.