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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BCRABL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)一同组成FKBP-RAPFRB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,Western blot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-ABL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。
BCR-ABL fusion protein is the basis of the pathogenesis of chronic myeloid leukemia (CML). Among them, BCR-ABL can only be located in the cytoplasm, can not translocate to the nucleus is the key factor in its pathogenesis. Therefore, translocation of BCRABL into the nucleus may be a potential method of treatment of CML. In this study, the recombinant adenovirus of HA-2FKBP-ABD (HF2A) and FLAG-3NLS-FRB * (FN3R) was constructed by gene recombination technology. Together with rapamycin analog, the FKBP-RAPFRB system was constructed to transport K562 BCR-ABL oncoproteins in the cytoplasm of the cells to the nucleus and explore its effect on the proliferation of K562 cells. The results showed that the recombinant adenovirus with high titer was successfully constructed and the target protein was successfully expressed in K562 cells by Western blot. The FKBP-RAP-FRB system inhibits K562 cell growth and clonogenicity by translocating BCR-ABL into the nucleus. The results revealed that translocation of BCR-ABL into the nucleus by the FKBP-RAP-FRB system is expected to provide a new therapeutic approach for CML.