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目的运用siRNA抑制胃癌细胞株SGC7901中NF-κB p65基因的表达,探讨其对细胞增殖和凋亡的影响,并分析该基因对化疗药物5-氟尿嘧啶(5-Fu)耐药性的作用。方法将SGC7901细胞分为空白对照组、脂质体组、5-Fu组、siRNA组和联合组,除空白对照组外,各组分别给予相应的药物。Western blot法检测各组细胞NF-κBp65蛋白的表达;Annexin V-PI法检测细胞凋亡;MTT法检测各组细胞的增殖水平。结果NF-κBp65蛋白在空白对照组和脂质体组中高表达,在5-Fu组、siRNA组和联合组中低表达,其中以联合组表达量最低;各组细胞早期凋亡率分别为1.40%±0.20%、2.36%±0.58%、6.68%±0.34%、6.60%±0.64%和21.62%±1.12%,其中以联合组最高;细胞的增殖活性随着siRNA浓度的增加和作用时间的延长而下降。结论siRNA能有效抑制NF-κBp65基因的表达,从而抑制SGC7901细胞增殖,促进其凋亡,并增强肿瘤细胞对化疗药物的敏感性。
Objective To investigate the effect of siRNA on the expression of NF-κB p65 gene in gastric cancer cell line SGC7901 and the effect of siRNA on 5-Fu resistance. Methods SGC7901 cells were divided into blank control group, liposome group, 5-Fu group, siRNA group and combined group, except the blank control group, each group were given the corresponding drugs. Western blot was used to detect the expression of NF-κBp65 protein in each group; Annexin V-PI was used to detect the apoptosis; MTT method was used to detect the proliferation of each group. Results The NF-κBp65 protein was highly expressed in the blank control group and the liposome group, but was low in the 5-Fu group, siRNA group and the combination group, and the lowest expression in the combination group. The early apoptosis rates in each group were 1.40 % ± 0.20%, 2.36% ± 0.58%, 6.68% ± 0.34%, 6.60% ± 0.64% and 21.62% ± 1.12%, respectively, which were the highest in the combination group. The proliferation activity of cells increased with the increase of siRNA concentration and the extension of the action time And down. Conclusion siRNA can effectively inhibit the expression of NF-κBp65, thereby inhibiting the proliferation and promoting the apoptosis of SGC7901 cells and enhancing the sensitivity of tumor cells to chemotherapeutic drugs.