目的探讨沉默细胞外信号调节激酶5(extracellular signal regulated kinase 5,ERK5)对胃癌细胞SGC-7901、BGC-823生物学功能的影响。方法实时荧光定量PCR法(qRT-PCR)检测人胃癌细胞株和人胃黏膜上皮细胞株ERK5mRNA的表达水平。应用短发...目的探讨沉默细胞外信号调节激酶5(extracellular signal regulated kinase 5,ERK5)对胃癌细胞SGC-7901、BGC-823生物学功能的影响。方法实时荧光定量PCR法(qRT-PCR)检测人胃癌细胞株和人胃黏膜上皮细胞株ERK5mRNA的表达水平。应用短发荚RNA(shRNA)干扰技术沉默胃癌细胞SGC-7901、BGC-823中ERK5的表达。CCK-8法检测ERK5沉默后胃癌细胞的生长能力,平板克隆实验检测细胞克隆形成能力,Transwell实验检测细胞侵袭和迁移能力,流式细胞仪检测细胞凋亡和周期分布。结果与胃黏膜上皮细胞GES-1相比,胃癌细胞SGC-7901、BGC-823、AGS、HGC-27中ERK5 mRNA均呈高表达,相对表达量分别为2.696±0.501、1.865±0.185、1.793±0.137和1.530±0.093(P<0.05)。ERK5-shRNA有效沉默胃癌细胞SGC-7901、BGC-823中ERK5的表达水平,沉默效率分别为(74.4±1.5)%和(69.1±3.9)%,差异有统计学意义(P<0.05)。沉默ERK5的表达能有效抑制胃癌细胞增殖、克隆形成、迁移和侵袭能力(P<0.05),而促进胃癌细胞凋亡(P<0.05),并诱导细胞周期阻滞于G0/G1期。结论沉默ERK5可显著抑制胃癌细胞SGC-7901、BGC-823的生长侵袭,促进细胞凋亡,ERK5可能是胃癌治疗的潜在靶点。展开更多
AIM: To explore the effect of trichostatin A (TSA) on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS: The effect of TSA on growth inhibition and apoptosis was exa...AIM: To explore the effect of trichostatin A (TSA) on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS: The effect of TSA on growth inhibition and apoptosis was examined by MTT, fluorescence microscopy and PI single-labeled flow cytometry. The acetylated histone H3 level was detected by Western blot. RESULTS: TSA induced apoptosis in gastric cancer cell lines BGC-823 and SGC-7901 was in a dose and time-dependent manner. Apoptotic cells varied significantly between TSA treated groups (37.5 ng/mL 72 h for BGC-823 cell line and 75 ng/mL 72 h for SGC-7901 cell line) and control group (0.85 ± 0.14 vs 1.14 ± 0.07, P = 0.02; 0.94 ± 0.07 vs 1.15 ± 0.06, P = 0.02). Morphologic changes of apoptosis, including nuclear chromatin condensation and fluorescence strength, were observed under fluorescence microscopy. TSA treatment in BGC-823 and SGC-7901 cell lines obviously induced cell apoptosis, which was demonstrated by the increased percentage of sub-G1 phase cells, the reduction of G1-phase cells and the increase of apoptosis rates in flow cytometric analysis. The result of Western blot showed that the expression of acetylated histone H3 increased in BGC-823 and SGC-7901 TSA treatment groups as compared with the control group.CONCLUSION: TSA can induce cell apoptosis in BGC-823 and SGC-7901 cell lines. The expression of acetylated histone H3 might be correlated with apoptosis.展开更多
文摘AIM: To explore the effect of trichostatin A (TSA) on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS: The effect of TSA on growth inhibition and apoptosis was examined by MTT, fluorescence microscopy and PI single-labeled flow cytometry. The acetylated histone H3 level was detected by Western blot. RESULTS: TSA induced apoptosis in gastric cancer cell lines BGC-823 and SGC-7901 was in a dose and time-dependent manner. Apoptotic cells varied significantly between TSA treated groups (37.5 ng/mL 72 h for BGC-823 cell line and 75 ng/mL 72 h for SGC-7901 cell line) and control group (0.85 ± 0.14 vs 1.14 ± 0.07, P = 0.02; 0.94 ± 0.07 vs 1.15 ± 0.06, P = 0.02). Morphologic changes of apoptosis, including nuclear chromatin condensation and fluorescence strength, were observed under fluorescence microscopy. TSA treatment in BGC-823 and SGC-7901 cell lines obviously induced cell apoptosis, which was demonstrated by the increased percentage of sub-G1 phase cells, the reduction of G1-phase cells and the increase of apoptosis rates in flow cytometric analysis. The result of Western blot showed that the expression of acetylated histone H3 increased in BGC-823 and SGC-7901 TSA treatment groups as compared with the control group.CONCLUSION: TSA can induce cell apoptosis in BGC-823 and SGC-7901 cell lines. The expression of acetylated histone H3 might be correlated with apoptosis.
