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目的探讨干细胞因子(SCF)/c-KIT系统对Bx PC-3胰腺癌细胞侵袭能力的影响以及低氧诱导因子1α(HIF-1α)的作用。方法分别在正常氧分压条件下加入SCF以及低氧环境培养Bx PC-3细胞,使用实时定量PCR和Western blot法检测HIF-1αmRNA和蛋白水平的变化;设计并合成HIF-1α特异性的小干涉RNA(si HIF-1α),转染Bx PC-3细胞,实时定量PCR和Western blot法分别检测SCF和下调HIF-1α水平后对基质金属蛋白酶2(MMP2)、MMP9和尿激酶型纤溶酶原激活物(u PA)表达的影响,TranswellTM检测SCF和下调HIF-1α水平后对Bx PC-3细胞侵袭能力的影响。结果 SCF和低氧刺激均可以上调HIF-1α的蛋白表达;si HIF-1α可以高效、特异性抑制Bx PC-3细胞中HIF-1α的蛋白表达;SCF能够促进MMP2、MMP9和u PA的表达,而敲低HIF-1α后下调上述基因的表达;SCF作用后Bx PC-3细胞的侵袭能力增强,而敲低HIF-1α后促进Bx PC-3细胞的侵袭。结论 SCF/c-KIT系统可以通过HIF-1α上调MMP2、MMP9和u PA的表达,从而增强Bx PC-3细胞的侵袭能力,而敲低HIF-1α可抑制其作用。
Objective To investigate the effect of stem cell factor (SCF) / c-KIT system on invasiveness of Bx PC-3 pancreatic cancer cells and the role of hypoxia inducible factor 1α (HIF-1α). Methods Bx PC-3 cells were cultured in SCF under hypoxic conditions and hypoxia condition respectively. The mRNA and protein levels of HIF-1α were detected by real-time PCR and Western blot. The specificity and specificity of HIF-1α The expression of MMP-2, MMP-9 and urokinase-type fibrinolysis in SCF and HIF-1alpha were detected by real-time quantitative PCR and Western blot after transfection of si HIF-1alpha, Effect of TranswellTM on Invasion Ability of Bx PC-3 Cells after SCF and HIF-1α Down-regulated. Results Both HIF-1α and HIF-1α were up-regulated by both SCF and hypoxia. HIF-1α inhibited the expression of HIF-1α in Bx PC-3 cells efficiently and specifically. SCF promoted the expression of MMP2, MMP9 and u PA , And down-regulated the expression of HIF-1α. The invasion ability of Bx PC-3 cells was enhanced after SCF treatment, but knockdown of HIF-1α promoted the invasion of Bx PC-3 cells. Conclusions SCF / c-KIT system can up-regulate the expression of MMP2, MMP9 and u PA by HIF-1α to enhance the invasiveness of Bx PC-3 cells. However, knockdown of HIF-1α can inhibit its effect.