反义PCNA与反义bcl-2脱氧寡核苷酸抑制人视网膜色素上皮生长

Inhibition of antisense PCNA and antisense bcl 2 oligodeoxynucleotides cultured human retinal pigment epithelial cells

目的 观察反义增生细胞核抗原 (PCNA)和反义 bcl-2脱氧寡核苷酸对培养的人视网膜色素上皮细胞 (RPE)生长的作用 .方法 用 DNA合成仪合成有 1 8个核苷酸的反义PCNA和 1 5个核苷酸的反义 bcl- 2脱氧寡核苷酸片段 ,以不同浓度加入 RPE培养液 ,培养 2～ 4d,用 SABC法检测细胞PCNA和 bcl- 2的表达 ,用 MTT法及流式细胞仪测定细胞抑制率 .结果 6 .2 5～ 1 0 0μm ol· L- 1 的反义 PCNA对 RPE的抑制率为 8.3%～ 38.4% ,呈剂量依赖性 ,并抑制细胞 PCNA的表达 .反义 bcl- 2未显示抑制作用 .结论 反义 PCNA脱氧寡核苷酸可抑制近 40 %的 RPE生长 。

AIM To observe the effects of antisense PCNA (proliferating cell nuclear antigen) and antisense bcl 2 oligodeoxynucleotides (ODN) on the growth of cultured human retinal pigment epithelial cells (RPE). METHODS Antisense PCNA ODN fragments including 18 mer and 15 mer antisense bcl 2 ODN were synthesized with a DNA synthesizer. Cell cultures were added with the antisense ODNs and maintained for 24 h to 48 h. The rates of cell inhibition were measured by MTT and flow cytometry, and the expression of PCNA and bcl 2 was measured by SABC techniques. RESULTS The inhibition rates of antisense PCNA ODN from 6.25 μmol·L -1 to 100 μmol·L -1 were 8.3% to 38 4% in a dose dependent manner and inhibited the expression of PCNA. Antisense bcl 2 ODN failed to show any inhibitory effects on cell growth. CONCLUSION Antisense PCNA ODN can inhibit up to 40% of RPE growth. and further study is needed for its clinical application of the antisense ODNs.