反基因及反义寡核苷酸体外抗前列腺癌作用的研究  

The effects of anti-gene and antisense oligonucleotides on prostate cancer in vitro

作　　者：张勇[1] 陈维真[1] 谢瑶[2] 姚志彬[2] 温星桥[3] 

机构地区：[1]中山大学附属第三医院核医学科,广州510630 [2]中山大学基础医学院解剖学教研室 [3]中山大学附属第三医院泌尿外科,广州510630

出　　处：《中华泌尿外科杂志》2005年第7期472-475,共4页Chinese Journal of Urology

基　　金：广东省自然科学基金资助项目(2001738);广东省医学科学基金资助项目(B2000048)

摘　　要：目的探讨三螺旋形成寡核苷酸(TFO)和反义寡核苷酸(ASO)对前列腺癌细胞雄激素受体(AR)表达和细胞增殖的影响。方法设计并合成针对AR基因的TFO和ASO,经脂质体介导转染前列腺癌细胞株LNCaP。MTT法检测细胞增殖活性,RTPCR检测AR基因表达,免疫组化法检测AR蛋白表达,放射免疫分析检测PSA表达。结果转染后24、48、72h,TFO组LNCaP细胞ARmRNA平均表达水平分别为0.25、0.33、0.46,显著低于ASO组的0.44、0.54、0.60,P<0.05;TFO对LNCaP细胞的生长抑制率(51.8%、65.8%、36.2%)显著高于ASO(28.8%、42.6%、17.5%),P<0.05。TFO对AR蛋白表达的抑制作用强于ASO。转染后24hPSA表达水平[(0.5±0.1)ng/ml]显著低于ASO组[(0.8±0.2)ng/ml],P<0.05。结论在LNCaP细胞TFO对AR表达和癌细胞增殖的抑制作用明显强于ASO,反基因策略对于前列腺癌的治疗具有重要研究价值。Objective To investigate the effects of triple-helix forming oligonucleotides (TFO) and antisense oligonucleotides (ASO) on androgen receptor (AR) expression and proliferation of prostate cancer cells. Methods The TFO and ASO specific for AR gene were designed and synthesized to transfect LNCaP cells with lipofectin.The cellular proliferation was detected by MTT assay.The expression of AR gene and protein was examined by RT-PCR and immunohistochemical study. The PSA expression was examined by radioimmunoassay. Results At 24,48 and 72 h after transfection, the AR mRNA expression levels of TFO group (0.25,0.33,0.46,respectively) were markedly lower than those of ASO group (0.44,0.54, 0.60 ,respectively,P<0.05).The inhibitory rates of TFO (51.8%, 65.8%, 36.2%, respectively) for cellular proliferation was significantly higher than those of ASO (28.8%,42.6%,17.5%,respectively,P< 0.05 ).At 24 h after transfection, the PSA expression level of TFO group [(0.5±0.1)ng/ml] was also significantly lower than that of ASO group [(0.8±0.2)ng/ml,P<0.05]. Conclusions The inhibitory effect of the TFO on AR expression and cell proliferation of LNCaP cells is significantly higher than that of ASO.The anti-gene strategy has important value in the study of prostate cancer gene therapy.

关 键 词：反义寡核苷酸 抗前列腺癌 LNCAP细胞 三螺旋形成寡核苷酸 前列腺癌细胞株 免疫组化法检测 LNCAP细胞 体外 细胞增殖活性 MTT法检测 0.05 蛋白表达 抑制作用 雄激素受体 ASO TFO 脂质体介导 PCR检测 生长抑制率 癌细胞增殖 

分 类 号：R737.25[医药卫生—肿瘤]