目的 观察骨形成发生蛋白4(BMP4)在U251细胞中的作用.方法 阿霉素、BMP4质粒体分别给予U251细胞,BMP4的小干扰给予阿霉素处理过48h的U251细胞,逆转录-聚合酶链反应(RT-PCR)、荧光定量PCR和Western blot检测转染是否成功及BMP4的表达量.噻唑蓝(MTT)检测细胞活性,应用公式(1-处理组A/空白组A)×100%计算处理组细胞增长抑制率.结果 RT-PCR、荧光定量PCR和Western blot证明转染是成功的.在给予BMP4质粒体0.5、1.0、1.5、2.0、2.5μg/孔后,细胞的抑制率分别为(23.4±1.1)%、(54.8±1.3)%、(58.8±1.6)%、(57.2±1.4)%、(56.1±0.9)%(P〈0.05),0.5μg/孔与1.0、1.5、2.0、2.5 μg/孔的抑制率差异有统计学意义(P〈0.05),1.0、1.5、2.0、2.5μg/孔之间差异无统计学意义(P〉0.05).单用阿霉素对U251细胞的抑制率为(45.2±1.1)%(P〈0.05).给予阿霉素后给予BMP4的siRNA,细胞的抑制率为(23.1±2.7)%,与阿霉素组比较差异有统计学意义(P〈0.05).结论 BMP4可以抑制胶质瘤细胞U251的生长.
Objective To to investigate the role of bone morhogenetic proteins 4 (BMP4) in the human brain glioma cell line U251. Methods Adriamycin or plasmid of BMP4 was given to U251 cells, and BMP4 siRNA was given to U251 cells which had been given adriamycin for 48 h. Methyl thiazolyl tet-razolium ( MTT) was used to measure the activity of U251 cell growth in control group and treatment group. By the formula (1-Atreatment group/Acontrol group) ×100% , U251 cells growth inhibition rate was calculated. By using reverse transcription-polymerase chain reaction ( RT-PCR) , fluorescence quantitative PCR (FQ-PCR) and Western blotting, the transfection efficiency and the expression of BMP4 were assayed. Results Transfection was proved to be successful by RT-PCR, quantitative fluorescent PCR and Western blotting. Growth inhibition rate of U251 cells which were given BMP4 plasmid was (23.4± 1.1)%, (54.8±1.3)%, (58.8 ±1.6)%,(57.2 ±1.4)%, (56.1±0.9)% at the 0.5, 1.0, 1.5, 2. 0, 2. 5μg/well. There was significant difference in inhibition rate between 0. 5μg/well group and 1.0, 1.5, 2.0, 2.5μg/well groups (P〈0.05), but no significant difference was found among 1.0, 1.5, 2.0, 2.5μg/well groups (P〉0.05). After adriamycin was added to U251 cells, the inhibition rate was (45. 2 ± 1. 1) % (P 〈0. 05 ). After BMP4 siRNA was added to U251 cells which had been given adriamycin , the inhibition rate of cells was (23.1± 2.7) % (P〈0.05). Conclusion BMP4 can inhibit the glioma cells U251. BMP4 may be a new target for glioma therapy.