摘要
目的:研究乏氧及人乏氧诱导因子-1α(HIF-1α)基因表达对舌鳞癌Tca8113细胞系黏附和侵袭能力的影响。方法:将化学合成的siRNAHIF-1α转染入Tca8113细胞后进行常氧或乏氧(1%O2)培养。实验设以下对照组:空白对照组、脂质体组及非特异性siRNA(siRNAIrr)组。采用real time-PCR和Western blot法测定细胞中HIF-1αmRNA表达和蛋白含量,并分别检测HIF-1α基因干扰前后细胞对细胞外基质(ECM)的黏附与侵袭力。结果:乏氧能够诱导Tca8113细胞的黏附与侵袭力增加。乏氧培养条件下,HIF-1α的表达上调主要发生在蛋白水平。siRNAHIF-1α转染后常氧或乏氧培养24 h,HIF-1αmRNA表达显著下调,与各对照组相比有统计学意义(P<0.01),Tca8113细胞内HIF-1α蛋白含量亦显著降低。无论在常氧还是乏氧培养条件下,siRNAHIF-1α转染的Tca8113细胞黏附力与各对照组相比均显著降低(P<0.05或P<0.01);侵袭力也显著降低(P<0.01)。乏氧条件下siRNAHIF-1α转染的Tca8113细胞黏附力和侵袭力下降程度高于常氧培养〔(36.4±2.7)%vs(26±2.35);(44.2±2.2)%vs(35±1.75),P<0.01)〕。结论:化学合成的靶向HIF-1α的siRNA能够下调Tca8113细胞中HIF-1α基因的表达,降低细胞对ECM的黏附和侵袭力,可能成为抑制肿瘤转移的基因治疗的新途径或新靶点。
Objective： To evaluate the effect of synthesized small interfering RNA targeting to HIF-1α on the adhesion and invasion of human tongue squamous cell carcinoma cell line （Tca8113 ）. Methods： A double strand small interference RNA （siRNA） targeting HIF-1α （siRNAHIF-1α） was transfected into cultured Tca8113 ceils by lipofectamine 2000. The expression of HIF-1α was investigated on mRNA level by real time-PCR and protein level by Western blot. The adhesion and invasion of Tca8113 cells to extracellular matrix （ECM） was also analyzed. Results： Exposure to hypoxia induced a prolonged elevation of HIF-1α protein and siRNAHIF-1α reduced HIF-1α synthesis as measured on mRNA level and protein level compared with the controls. No matter under normoxic or hypoxic conditions, the adhesion potency of siRNAHIF-1α treated Tca8113 cells was markedly inhibited compared with controls（P 〈0.05 or P 〈 0.01 ）. So did the invasion potency （P 〈 0.01 ）. The adhesion and invasion potency of siRNAHIF-1α treated Tca8113 ceils were inhibited more greatly under hypoxic condition than under normoxic condition [ （ 36.4 ± 2.7 ） % vs （ 26 ± 2.35 ） ; （ 44.2 ± 2.2 ） % vs （35 ± 1.75）, P 〈0.01 ）]. Conclusion： siRNAHIF-1α can knockdown the expression of HIF-1α and inhibit the cell adhesion and in- vasion to ECM in Tca8113 cells. HIF-1α may play an established role in the regulation of Tca8113 cells invasion and metastasis. Interfering with HIF-1α pathways by siRNA strategy may provide a therapeutic target for human tongue squamous cell carcinomas.
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2009年第6期828-832,共5页
Journal of Practical Stomatology
基金
国家自然科学基金资助项目(编号:30572056)
