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目的:研究甲胎蛋白特异siRNA腺病毒载体对人甲胎蛋白阳性胃癌细胞FU97AFP基因的抑制作用,及抑制甲胎蛋白基因后对此肿瘤细胞周期及凋亡的影响。方法:利用构建好的腺病毒Ad-EGFP-U6-AFP-siRNA载体转染入FU97细胞,筛选出最佳感染复数。结果:筛选出最佳感染复数为:MOI=100;Ad-EGFP-U6-AFP-siRNA转染细胞后能显著抑制AFP基因在mRNA和蛋白水平的表达,G0/G1期细胞百分率〔(33.49±1.85)%〕明显低于空载体对照组和空白对照组〔(64.76±2.98)%和(74.81±2.60)%,P<0.01〕,G2/S期〔(64.10±2.59)%〕显著高于空载体组和空白对照组〔(34.77±1.91)%和(29.85±2.85)%〕,P<0.01;凋亡分析,实验组与各对照组之间凋亡发生差异无统计学意义,P>0.05。结论:重组腺病毒介导的RNA干扰能够显著抑制AFP在FU97中mRNA水平和蛋白水平的表达;抑制AFP后,影响细胞增殖周期,但对细胞凋亡无影响。
OBJECTIVE: To study the inhibitory effect of AFP-specific siRNA adenovirus vector on the gene expression of FU97AFP in human alpha-fetoprotein-positive gastric cancer cells and its effect on the cell cycle and apoptosis after the inhibition of alpha-fetoprotein gene. Methods: The constructed recombinant adenovirus Ad-EGFP-U6-AFP-siRNA vector was transfected into FU97 cells to screen out the best multiplicity of infection. Results: The optimal multiplicity of infection was selected as MOI = 100. The expression of AFP gene at mRNA and protein level was significantly inhibited by Ad-EGFP-U6-AFP-siRNA transfected cells. The percentage of cells in G0 / (64.76 ± 2.98)% and (74.81 ± 2.60)% respectively, P <0.01〕, G2 / S phase 〔(64.10 ± 2.59)%〕 was significantly higher than that in control group and blank control group There was no significant difference in apoptosis between experimental group and control group (P> 0.05), there was no significant difference between the control group and blank vector group (34.77 ± 1.91) and (29.85 ± 2.85)%, P < 0.05. CONCLUSION: Recombinant adenovirus-mediated RNA interference can significantly inhibit the expression of AFP mRNA and protein in FU97 cells. Inhibition of AFP can affect the cell cycle but has no effect on apoptosis.