RNA干扰抑制HMGB1基因表达对肺癌细胞L9981增殖和侵袭的影响

Effect on Invasion and Proliferation of Human Lung Cancer Cell L9981 by RNAi Inhibiting HMGB1 Gene Expression

背景与目的研究小干扰RNA抑制高迁移族蛋白B1基因表达对肺癌细胞L9981增值和侵袭能力的影响。方法设计合成靶向HMGB1基因的siRNA,采用阳离子脂质体试剂瞬时转染肺癌细胞株L9981,利用实时荧光定量PCR和Western blot检测RNA干扰后HMGB1基因的沉默效果,评价siRNA设计的合理性及RNA干扰抑制HMGB1表达的有效性;用细胞活力计数仪测定3组细胞活力;分别在RNA干扰后24、48、72和96小时应用MTT实验检测细胞生存状态,计算生长抑制率并绘制生长曲线;应用boyden chamber法检测侵袭能力的差异。结果靶向HMGB1的siRNA成功抑制肺癌细胞株L9981中HMGB1基因及蛋白表达,细胞活力检测仪测定RNA干扰48小时后,细胞活力显著下降;MTT测定肺癌细胞生长受到明显抑制;boyden chamber小室细胞侵袭实验结果肺癌穿膜细胞数明显减少。结论应用siRNA技术能有效的抑制HMGB1基因的表达,同时有效抑制L9981细胞的体外增值和侵袭,为肺癌的生物学治疗提供了新思路。

Background and objective To study the effects and mechanism of HMGB1 small interfering RNA （siRNA） on invasion and proliferation of human lung cancer cell. Methods After lung cancer cell line L9981 were transfected by HMGB 1 siRNA, the mRNA and protein of PRL21 were determined by real time PCR and western blot assay, respectively. The anchorage-independent growth was exmined by clonformation in soft agar, and invasion ability was evaluated by boyden chamber model. Results The siRNA could down-regulate the level of mRNA and protein of HMGB1. Suppression of of HMGB1 expression can inhibit invasion and proliferation ability in human lung cancer cell L9981. Conclusion HMGB1 gene might play an important role in development of human lung cancer, and down-regulation by HMGB1 siRNA could inhibit invasion and proliferation of human lung cancer cell.