论文部分内容阅读
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 effect of triple helix-forming oligonucleotide (TFO) and antisense oligonucleotide (ASO) on the expression of androgen receptor (AR) and cell proliferation in prostate cancer cells. Methods TFO and ASO targeting to AR gene were designed and synthesized and transfected into LNCaP prostate cancer cell line by liposome. Cell proliferation was detected by MTT assay, AR gene expression by RTPCR, AR protein by immunohistochemistry and PSA expression by radioimmunoassay. Results At 24, 48 and 72 h after transfection, the average mRNA expression of AR mRNA in LNCaP cells in TFO group was 0.25, 0.33 and 0.46, respectively, which was significantly lower than that in ASO group (0.44,0.54,0.60, P <0.05). The growth inhibition rate of LNCaP cells (51.8%, 65.8%, 36.2%) were significantly higher than ASO (28.8%, 42.6%, 17.5%), P <0.05. The inhibitory effect of TFO on AR protein expression was stronger than that of ASO. The expression ofPSA at 24h after transfection was significantly lower than that in ASO group [(0.5 ± 0.1) ng / ml] [(0.8 ± 0.2) ng / ml], P <0.05. Conclusion The inhibitory effect of TFO on AR expression and cancer cell proliferation in LNCaP cells is stronger than that in ASO. The anti-gene strategy is of great value in the treatment of prostate cancer.