HIF-1α基因沉默对人胰腺癌Patu8988细胞增殖和细胞周期的影响

Effects of RNA Interference Mediated Gene Silencing of Hypoxia Inducible Factor-1α on the Cell Proliferation and Cell Cycle of the Human Pancreatic Cancer Patu8988 Cells

目的:观察沉默HIF-1α基因对人胰腺癌Patu8988细胞增殖和细胞周期的影响。方法:构建针对乏氧诱导因子-1α(HIF-1α)基因的RNA干扰(RNAi)慢病毒表达载体LV-RNAi-HIF-1α并转染人胰腺癌Patu8988细胞,MTT法观察HIF-1α基因沉默后胰腺癌Patu8988细胞体外增殖情况,以空白组及转染阴性对照干扰载体的阴性组为对照;流式细胞仪检测Patu8988/LV-RNAi-HIF-1α的细胞周期,以空白组为对照。结果:与空白组及阴性组细胞相比,Patu8988/LV-RNAi-HIF-1α的细胞增殖速度减慢,差异有统计学意义(F=58.48,P=0.000;F=55.91,P=0.000);Patu8988/LV-RNAi-HIF-1 G0/G1细胞比例较空白组高(t=10.87,P=0.001),S期、G2/M期细胞比例较空白组低(t=6.44,P=0.005,t=5.49,P=0.005),表现出G0/G1期阻滞。结论:HIF-1α基因表达沉默抑制人胰腺癌Patu8988细胞的DNA合成,将细胞阻滞于G0/G1期,使Patu8988细胞增殖能力降低,胰腺癌细胞体外生长被显著抑制。

Objective To investigate the influence of RNA interference on expression of HIF-1α gene on the cell proliferation and cell cycle of the human pancreatic cancer cell line Patu8988. Methods Construet the lentiviral veetor-mediated RNA interference of HIF-1α Patu8988 cells transfected with the empty vector served as hypoxia negative control group, Patu8988 cells not transfeeted with vector served as hypoxia blank control group, cells eyele and cell proliferation were evaluated by using flow-cytometric anlysis and MTr assay. Results The growth of the pancreatic cancer cell was decreased obviously compared to the blank control group and negative control group （F = 58.58, P = 0. 000; F = 55.91, P = 0. 000） ; In addition, the percentage of cells in Go/G1 phase in experimental group was significantly more than that in blank control group （t = 10.87, P =0.01）, the pereentage of cells in S and G2/M phase in experimental group were less than that in blank control group （ t = 6.44, P = 0.05 ; t = 5.49,P = 0.005 ）. Conclusion Gene silencing of HIF-1α using RNAi lentiviral vector-mediated RNA can inhibit the expression of HIF-1α, inhibit the proliferation and cell cycle at G0/G1 phase, decrease the growth of the Patu8988 pancreatic cancer cells in vitro.