CAJ | 학술논문

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siRNA_(HIF-1α)转染入Tca8113细胞后进行常氧或乏氧(1% O_2)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(siRNA_(Irr))组.采用real time-PCR和Western blot法测定细胞中HIF-1α mRNA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.siRNA_(HIF-1α)转染后常氧或乏氧培养24 h, HIF-1α mRNA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,siRNA_(HIF-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.
목적:연구핍양급인핍양유도인자-1α(HIF-1α)기인표체대설린암Tca8113세포계점부화침습능력적영향.방법:장화학합성적siRNA_(HIF-1α)전염입Tca8113세포후진행상양혹핍양(1% O_2)배양.실험설이하대조조:공백대조조、지질체조급비특이성siRNA(siRNA_(Irr))조.채용real time-PCR화Western blot법측정세포중HIF-1α mRNA표체화단백함량,병분별검측HIF-1α기인간우전후세포대세포외기질(ECM)적점부여침습력.결과: 핍양능구유도Tca8113세포적점부여침습력증가.핍양배양조건하,HIF-1α적표체상조주요발생재단백수평.siRNA_(HIF-1α)전염후상양혹핍양배양24 h, HIF-1α mRNA표체현저하조,여각대조조상비유통계학의의(P<0.01),Tca8113세포내HIF-1α단백함량역현저강저.무론재상양환시핍양배양조건하,siRNA_(HIF-1α)전염적Tca8113세포점부력여각대조조상비균현저강저(P<0.05혹P<0.01);침습력야현저강저(P<0.01).핍양조건하siRNAHIF-1α전염적Tca8113세포점부력화침습력하강정도고우상양배양[(36.4±2.7)% vs (26±2.35);(44.2±2.2)% vs (35±1.75), P<0.01)].결론: 화학합성적파향HIF-1α적siRNA능구하조Tca8113세포중HIF-1α기인적표체,강저세포대ECM적점부화침습력,가능성위억제종류전이적기인치료적신도경혹신파점.
Objective: To evaluate the effect of synthesized small interfering RNA targeting to HIF-lα 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 (siRNAH1Fla) was transfected into cultured Tca8113 cells by lipofectamine2000. 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-lα protein and siRNAHIF.la reduced HIF-la synthesis as measured on mRNA level and protein level compared with the controls. No matter under normoxic or hy-poxic 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.,a treated Tca8113 cells 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; siRNAH1F.lo can knockdown the expression of HIF-la and inhibit the cell adhesion and invasion to ECM in Tca8113 cells. HIF-la 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.