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目的研究三氧化二砷(As_2O_3)对胃癌 SGC-7901细胞作用过程中细胞内核转录因子kB(NF-kB)的激活及重组腺病毒 Ad-IKBαM通过抑制 NF-kB 的活化增强 As_2O_3的诱导凋亡作用。方法培养胃癌 SGC-7901细胞,以非感染组及感染 Ad-IkBα组为对照,采用凝胶电泳迁移率实验(EMSA)及免疫组化法检测 As_2O_3处理后细胞核内 NF-kB 的激活情况,以及感染 Ad-IkBαM对 NF-kB活性的影响;四甲基偶氮唑盐(MTT)法、Hoechest 染色、原位末端标记(TUNEL)法分别检测感染Ad-IkBαM对 As_2O_3诱导细胞凋亡的影响。结果 EMSA 及免疫组化法显示 As_2O_3作用于胃癌细胞可使细胞内 NF-kB 激活,感染 Ad-IkBαM使 NF-kB 活性受到明显抑制;MTT 法证明,As_2O_3作用后,感染 Ad-IkBαM细胞的凋亡率(59.2±2.5)%较感染 Ad-IkBα组(47.5±2.3)%及未感染组(40.0±1.2)%明显升高,各组间比较 P<0.01;Hoechest 法显示,感染 Ad-IKBαM组的凋亡率为(27.7±2.6)%,明显高于感染 Ad-IkBα组(18.3±1.5)%及未感染组(11.0±1.7)%(P<0.05)。TUNEL 法结果与 Hoechest 法一致,感染 Ad-IkBαM组的凋亡率为(31.1±2.5)%,高于感染 Ad-IkBα组(20.7±2.1)%及未感染组(13.0±1.7)%(P<0.05)。可见,感染 Ad-IkBαM可明显提高 As_2O_3诱导的细胞凋亡。结论 As_2O_3作用于胃癌细胞可使细胞内 NF-kB 激活,从而表明 NF-kB 激活可能为胃癌细胞抗诱导凋亡作用的重要机制;感染 Ad-IKBαM可有效抑制 NF-kB 的活性,并增强 As_2O_3的诱导凋亡作用。
Objective To investigate the effect of As 2 O 3 on the activation of intracellular nuclear factor kB (NF-kB) and the effect of recombinant adenovirus Ad-IKBαM on As 2 O 3 -induced apoptosis in gastric cancer SGC-7901 cells by inhibiting the activation of NF-κB. Methods SGC-7901 cells were cultured and the activation of NF-κB in As2 O3-treated SGC-7901 cells was detected by EMSA and immunohistochemistry in non-infected and Ad-IkBα infected groups The effect of Ad-IkBαM infection on the activity of NF-κB was detected by MTT assay, Hoechst staining and TUNEL method. Results EMSA and immunohistochemistry showed that As_2O_3 could activate NF-κB in gastric cancer cells. Ad-IkBαM inhibited the activity of NF-κB in gastric cancer cells. MTT assay showed that As_2O_3 induced apoptosis of Ad-IkBαM cells Compared with Ad-IkBα group (47.5 ± 2.3)% and uninfected group (40.0 ± 1.2)%, the mortality rate (59.2 ± 2.5)% was significantly higher than that in Ad-IkBα group (27.7 ± 2.6)%, which was significantly higher than that in Ad-IkBα group (18.3 ± 1.5%) and non-infected group (11.0 ± 1.7%) (P <0.05). TUNEL method was consistent with Hoechest method. The apoptosis rate in Ad-IkBαM group was (31.1 ± 2.5)%, higher than that in Ad-IkBα group (20.7 ± 2.1%) and uninfected group (13.0 ± 1.7%) <0.05). It can be seen that infection with Ad-IkBαM can significantly increase As 2 O 3 -induced apoptosis. Conclusion As 2 O 3 can activate NF-κB in gastric cancer cells, indicating that NF-κB activation may be an important mechanism of anti-apoptosis in gastric cancer cells. Infection with Ad-IKBαM can inhibit the activity of NF-κB and enhance the activity of As 2 O 3 Induced apoptosis.