雄激素受体反义RNA对前列腺癌细胞雄激素受体表达的抑制作用研究

目的 观测雄激素受体（AR）反义RNA逆转录病毒表达载体在前列腺癌细胞中的表达及其对AR表达的影响。方法 将AR反义RNA逆转录病毒表达载体导入前列腺癌细胞株LNCaP，PCR检测AR目的片段在重组LNCaP细胞基因组中的存在，RT－PCR检测AR反义RNA表达和AR mRNA水平，Western blot检测AR蛋白表达水平。结果 PCR在LNCaP^as-AR基因组DNA中扩增出了AR目的片段，表明AR目的片段整合到了细胞基因组。RT－PCR证实LNCaP^as-AR有AR反义RNA表达，且其AR mRNA水平较LNCaP和空载体转染的LNCaP^Neo显著下调（P＜0.05)，Western blot证实LNCaP^as-AR细胞AR蛋白含量显著下降（P＜0.05)。结论 AR反义RNA可抑制前列腺癌细胞AR表达。

A study of the inhibitive effect of androgen receptor antisense RNA on androgen receptor in prostate cancer cells

Objective To detect the expression of androgen receptor (AR) antisense RNA retroviral vector in prostate cancer cell LNCaP and the effect of AR antisense RNA on the expression of ARmRNA and its protein. Methods The AR antisense RNA retroviral vector was transfected into the prostate cancer cell LNCaP. The presence of the AR inserted fragment was examined by PCR in the genomic DNA of recombinant LNCaPas AR. The expression of AR antisense RNA and the level of AR mRNA were detected by RT PCR. The level of AR protein was detected by Western blot. Results A fragment containing the AR inserted fragment was amplified by PCR in the genomic DNA of LNCaPas-AR cells, demonstrating that the AR cDNA fragment was integrated into the genomic DNA of the cells. AR antisense RNA was detected in LNCaPas AR cells by RT PCR and the level of AR mRNA was significantly down regulated in LNCaPas AR as compared with LNCaP and LNCaPNeo ( P <0.05). Western blot analysis showed a significant reduction of AR protein in LNCaPas AR as compared with those of LNCaP and LNCaPNeo ( P <0.05). Conclusion AR antisense RNA can inhibit the AR expression in prostate cancer cell LNCaP.