下调人高迁移率族蛋白1对人前列腺癌细胞PC-3的影响

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。

Impact on human prostatic carcinoma cell PC-3 from down-regulation of HMGB1

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.MethodsTo 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 transfection 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.