摘要
目的探讨Moeisn蛋白参与影响人脑胶质细胞瘤(U251)细胞生长和侵袭的可能途径。方法以人脑胶质细胞瘤U251细胞系为研究对象,针对Moesin编码基因设计小干扰RNA片段转染入细胞,通过反转录聚合酶链反应(RT-PCR)筛选沉默效率最高的siRNA片段进行后续实验。通过对设计的3组不同干预的U251细胞(空白组、阴性对照组、转染组)进行RT-PCR测定PDGF的表达,以及流式细胞方法测定CD44的含量。结果 RT-PCR和Western-blot筛选出沉默效果最好的Moesin-139片段;RT-PCR发现转染组U251细胞中PDGF的mRNA的平均表达为0.09377±0.008546,远低于空白组(0.4663±0.01844)和阴性对照组(0.4570±0.02159),其差异有统计学意义(P<0.01);流式细胞术检测发现转染组中CD44的表达最低为(26.73±2.720)%,与其他两组差异明显(F=173.669,sig=0.000)。结论 U251细胞中Moesin表达下降将导致PDGF和CD44的表达同时下降,同时生长和侵袭能力明显下降,提示Moesin可能通过调节PDGF与CD44在人脑胶质瘤细胞的生长、侵袭过程中发挥重要作用,Moesin有可能成为治疗人脑胶质瘤的靶点分子。
Objective To explore the possible ways how Moeisn proteins involved in the growth and invasion of human brain glioblastoma（U251）.Methods The human brain glioblastoma cell line U251 was chosen as the research object.Small interfering RNA fragment was designed for Moesin coding gene and transfected into cells.Through the reverse transcription polymerase chain reaction（RT-PCR） was screen out the most efficient siRNA silencing fragments for follow-up experiments.Three different intervention U251 cells（blank control group,negative control and transfection group） were designed for RT-PCR determination of expression of PDGF,and the amount of CD44 was measured by flow cytometry.Results Screening of a fragment of Moesin-139 by RT-PCR and Western-blot was the best silent;RT-PCR found that PDGF mRNA expression of U251 cells in the transfected group was 0.09377±0.008546 on average which was much lower than that of the blank group（0.4663±0.01844） and negative control group was（0.4570±0.02159）.The difference was of obviously statistical significance（P0.01）.Flow cytometry detected that the expression of CD44 in transfected group was（26.73±2.720）% at minimum which had obvious differences with 2 other groups（F=173.669,sig=0.000）.Conclusion In U251 cells,the downregulation of the expression of Moesin will lead to PDGF and CD44 to decrease at the same time.Moesin probably plays an important role in the invasion and metastasis of in human glioma cell by regulating PDGF and CD44.Moesin may become a target molecule in the treatment of human glioma.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2011年第2期121-125,共5页
Cancer Research on Prevention and Treatment
基金
湖南省科技厅胶质瘤基金资助项目(JT3049)
