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目的将前列腺特异性启动子驱动的自杀基因导入到雄激素非依赖性前列腺癌(androgen independent prostate cancer,AIPC)细胞,观察其对AIPC细胞的杀伤作用。方法将rPB-DR(6)启动子驱动的胞嘧啶脱氨酶(CD)基因融合分子插入到逆转录病毒载体pLNCX,构建pLN-rPB-DR(6)-CD重组逆转录病毒载体,PA317包装,获得重组逆转录病毒,后者感染前列腺癌细胞PC-3,获得G418抗性细胞PC-3CD。在培养基中加入维甲酸、5-氟胞嘧啶,观察其对PC-3CD的杀伤作用。结果经酶切和测序证实成功构建了pLN-rPB-DR(6)-CD重组逆转录病毒载体,PCR证实在PC-3CD基因组DNA中扩增出了CD基因,RT-PCR证实PC-3CD有CD mRNA表达。在培养基中加入维甲酸、5-氟胞嘧啶后,PC-3CD细胞大量死亡,生长显著被抑制。结论前列腺特异性启动子驱动的自杀基因可杀伤雄激素非依赖性前列腺癌细胞,并显著抑制其生长。
Objective To induce prostate cancer-specific suicide gene into androgen independent prostate cancer (AIPC) cells and to observe its killing effect on AIPC cells. Methods The fusion gene of cytosine deaminase (CD) driven by rPB-DR (6) promoter was inserted into retrovirus vector pLNCX to construct pLN-rPB-DR (6) -CD recombinant retroviral vector and PA317 , Get recombinant retrovirus, the latter infected with prostate cancer cells PC-3, to obtain G418 resistant cells PC-3CD. Add retinoic acid, 5-fluorocytosine to the medium to observe the killing effect on PC-3CD. Results The recombinant retroviral vector pLN-rPB-DR (6) -CD was confirmed by restriction enzyme digestion and sequencing. PCR confirmed that the CD gene was amplified from PC-3CD genomic DNA. RT-PCR confirmed that PC-3CD CD mRNA expression. After adding retinoic acid and 5-fluorocytosine to the medium, PC-3CD cells died in large numbers and their growth was significantly inhibited. Conclusion Prostate-specific promoter-driven suicide genes can kill androgen-independent prostate cancer cells and significantly inhibit their growth.