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目的探讨环氧化酶-2(COX-2)短发夹RNA(shRNA)对胃癌细胞SGC-7901的生长和细胞周期的影响。方法构建COX-2基因的特异性小RNA干扰质粒,构建靶向抑制COX-2基因的重组表达载体pshRNA-COX-2。实验分3组:未转染胃癌细胞组,阴性对照HK组,pshRNA-COX-2转染组。用质脂体lipofectamineTM2000转染胃癌细胞SGC-7901,RT-PCR和Western blot分别检测COX-2基因mRNA和蛋白表达情况,流式细胞术检测细胞周期,细胞计数检测细胞的生长变化。结果与阴性对照组相比,pshRNA-COX-2重组表达载体抑制COX-2 mRNA及蛋白表达的抑制率分别为70.1%和43.2%;G0~G1期细胞由61.5%上升至70.2%,S期细胞由27.3%下降至21.7%;细胞生长明显减慢。结论pshRNA-COX-2重组表达载体能显著抑制COX-2的表达,从而抑制胃癌细胞生长。
Objective To investigate the effect of COX-2 short hairpin RNA (shRNA) on the growth and cell cycle of gastric cancer cell line SGC-7901. METHODS: The specific small interfering RNA (siRNA) plasmid of COX-2 gene was constructed and the recombinant expression vector pshRNA-COX-2 targeting COX-2 gene was constructed. The experiment was divided into 3 groups: untransfected gastric cancer cell group, negative control HK group, pshRNA-COX-2 transfection group. Gastric cancer cells SGC-7901 were transfected with lipofectamineTM2000. The expression of COX-2 mRNA and protein were detected by RT-PCR and Western blot respectively. The cell cycle and cell counts were detected by flow cytometry. Results Compared with negative control group, the inhibitory rates of pshRNA-COX-2 recombinant expression vector on COX-2 mRNA and protein expression were 70.1% and 43.2%, respectively. The percentage of cells in G0-G1 phase increased from 61.5% to 70.2% Cells decreased from 27.3% to 21.7%; cell growth slowed significantly. Conclusion pshRNA-COX-2 recombinant expression vector can significantly inhibit the expression of COX-2, thereby inhibiting the growth of gastric cancer cells.