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作 者:宋登新[1] 陈安民[1] 郭风劲[1] 谢柏臻[1] 廖晖[1] 朱波[1] 陈超[1]
机构地区:[1]华中科技大学同济医学院同济医院骨科,湖北武汉430030
出 处:《基础医学与临床》2008年第5期441-445,共5页Basic and Clinical Medicine
基 金:国家973计划基金(2002CB513100)
摘 要:目的研究小干扰RNA(siRNA)抑制高迁移率族蛋白1(HMGB1)基因对PC-3侵袭和转移的影响。方法构建真核表达载体Pgenesil-1/HMGB1siRNA,转染PC-3细胞系,通过RT-PCR和Western blot检测HMGB1的mRNA及蛋白;通过细胞划痕、transwell和明胶酶谱分析检测PC-3体外侵袭能力。结果成功构建siRNA表达载体Pgene-sil-1/HMGB1siRNA,可使PC-3细胞HMGB1mRNA和蛋白表达水平显著降低(P<0.05),并能有效抑制PC-3体外侵袭和转移活性(P<0.05)。结论应用siRNA技术能有效的抑制基因的表达,同时也能有效抑制PC-3细胞的体外侵袭和转移,为肿瘤的生物学治疗提供了新思路。Objective To study the inhibition effect of small interfering RNAs (siRNA) on High Mobility Group Box-1 gene expression in PC-3 cells, and to study the inhibitory effect on invasion and metabasis of PC-3. Methods To constructan eukaryotic expression vector carrying Pgenesil-1/HMGB1 siRNA ,then the vector was transfected into PC-3 and the positive colonies were screened by G418. HMGB1 expression of PC-3 cells before and after transfec- tion was detected by RT-PCR and Western blot. The migration and invasion were examined by scratch method, transwell and gel zymography. Results The recombinant plasmid Pgenesil-1/HMGB1 siRNA was successfully constructed. The introduction of Pgenesil-1/HMGB1 siRNA efficiently and specifically inhibited the expression of HMGB1 gene as shown by the results of RT-PCR and Western blot. There was a significant difference between the siRNA transfected group and the control groups ( P 〈 0. 05 ). The migration and invasion in Pgenesil-1/HMGB1 siRNA group was significantly lower than that the control groups ( P 〈 0. 05 ). Conclusion Pgenesil-1/HMGB1 siRNA can specially suppress the expression of HMGB1 gene in PC-3 cells, effectively inhibit the migration and invasion of PC-3. So it provides novo approach to bio-therapy of cancer.
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