论文部分内容阅读
目的 了解反义c myc基因稳定整合对正常细胞的影响。 方法 将分别载有c myc第 1、2和 3外显子的反义片段的 3种反义c myc重组逆转录病毒表达载体aM1、aM2和aM3,导入c myc正常表达的正常小鼠成纤维细胞系NIH3T3,并在基因稳定表达后进行各项检测。 结果 3种反义载体导入明显抑制了NIH3T3c myc表达(抑制作用aM2 >aM3>aM1)、PCNA表达 (aM2 >aM1>aM3) ,使细胞体外生长增殖活性下降 (下降程度aM2 >aM1>aM3) ,aM2、aM3还使细胞周期出现了延长和DNA合成减少 ,并使NIH3T3软琼脂集落形成能力下降 (aM2 >aM3)。 结论 反义c mycRNA的导入对正常细胞的生长增殖能力有抑制作用 ,提示局部治疗和靶向治疗在反义c myc基因治疗中的必要性
Objective To understand the effect of stable integration of antisense c myc gene on normal cells. Methods Three antisense c myc recombinant retroviral vectors aM1, aM2 and aM3 containing the antisense fragments of exons 1, 2 and 3, respectively, of c myc were introduced into normal mouse fibroblasts expressing c myc Cell line NIH3T3, and the gene is stable expression after the test. Results The expression of NIH3T3c myc (aM2> aM3> aM1) and PCNA (aM2> aM1> aM3) inhibited the proliferation and proliferation of NIH3T3c myc in vitro. aM2, aM3 also led to prolonged cell cycle and reduced DNA synthesis, as well as reduced NIH3T3 soft agar colony forming ability (aM2> aM3). Conclusion The introduction of antisense c mycRNA can inhibit the growth and proliferation of normal cells, suggesting the necessity of local therapy and targeted therapy in antisense c myc gene therapy