李天煜, 陈利生. 曲古抑菌素A 对人结肠癌细胞株Colo205 组蛋白乙酰化及ING1b mRNA表达的影响*[J]. 中国肿瘤临床, 2010, 37(4): 181-183. DOI: 10.3969/j.issn.1000-8179.2010.04.001
引用本文: 李天煜, 陈利生. 曲古抑菌素A 对人结肠癌细胞株Colo205 组蛋白乙酰化及ING1b mRNA表达的影响*[J]. 中国肿瘤临床, 2010, 37(4): 181-183. DOI: 10.3969/j.issn.1000-8179.2010.04.001
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LI Tianyu, CHEN Lisheng. The Effect of Trichostatin A on Histone Acetylation and mRNA Expression of ING1b Genein Human Colon Cancer Cell Line Colo 205[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(4): 181-183. DOI: 10.3969/j.issn.1000-8179.2010.04.001
Citation: LI Tianyu, CHEN Lisheng. The Effect of Trichostatin A on Histone Acetylation and mRNA Expression of ING1b Genein Human Colon Cancer Cell Line Colo 205[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(4): 181-183. DOI: 10.3969/j.issn.1000-8179.2010.04.001
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曲古抑菌素A 对人结肠癌细胞株Colo205 组蛋白乙酰化及ING1b mRNA表达的影响*

The Effect of Trichostatin A on Histone Acetylation and mRNA Expression of ING1b Genein Human Colon Cancer Cell Line Colo 205

    摘要
  • 摘要: 目的：探讨曲古抑菌素A（TSA ）对人结肠癌细胞株Colo 205 组蛋白乙酰化及ING 1b mRNA 表达的影响。方法：培养人结肠癌细胞株Colo 205，对照组（A 组）不加TSA 干预，实验组分3 组（B、C、D 组），分别应用组蛋白去乙酰化酶（HDACs）抑制剂TSA50、100、200 μ g/L 的浓度作用于人结肠癌细胞株Colo 205，24h 后用染色质免疫沉淀（ChIP）方法检测4 组Colo 205 细胞乙酰化组蛋白H3 结合的DNA情况，以了解抑癌基因ING 1b 相关组蛋白H3 乙酰化的变化，并用逆转录聚合酶链反应（RT-PCR）方法检测ING 1b mRNA 的表达，均用实时定量PCR 方法分析。结果：A 组人结肠癌细胞株Colo 205 组蛋白H3 乙酰化水平及ING 1b mRNA Ct值为23.25± 0.08和23.32± 0.05，经TSA 干预后，C、D 组组蛋白H3 乙酰化水平较A 组增加（P<0.05），2-Δ Δ Ct值分别为4.21和4.38，ING 1b mRNA 表达亦比A 组高（P<0.05），2-Δ Δ Ct值分别为4.52和4.62，组蛋白H3 乙酰化水平及ING 1b mRNA 表达C、D 组间无显著性差异（P>0.05），组蛋白H3 乙酰化水平及ING 1b mRNA 表达B 组与A 组比较无显著性差异（P>0.05），2-Δ Δ Ct值分别为1.12和1.33。同时观察到C、D 组Colo 205 细胞较A、B 组细胞生长明显受抑制。结论：人结肠癌细胞株Colo 205 组蛋白去乙酰化可能是导致基因ING 1b 表达沉默的主要原因之一，100 μ g/L 的TSA 能较好地提高组蛋白乙酰化水平，并有效地激活去乙酰化所致ING 1b 基因转录，诱导该基因表达，从而抑制肿瘤细胞生长。

     

    Abstract: Objective:To investigate the effect of Trichostatin A (TSA) on histone acetylation and expression of ING1b mRNA in Colo205 human colon cancer cell line. Methods: Human colon cancer Colo205 cells were cultured and divided into 4 groups. Cells in the control group (group A) was treated without TSA. In the other three groups, cells were treated with 3 different concentrations of TSA:50μ g/L (group B), 100 μ g/L (group C), and 200 μ g/L (group D). At 24hours after treatment, the level of histone H3 acetylation was analyzed by chromatin immunoprecipitation (ChIP) and the expression of ING 1b mRNA was detected by RT-PCR with qPCR. The growth of Colo 205 human colon cancer cells in group C and D was obviously inhibited compared with that in group A and B.Results:The Ct value of histone H 3 acetylation and mRNA expression of ING1b in group A were 23.25±0.08and 23.32±0.05, respectively. After treatment with TSA, the 2-Δ Δ Ct value of histone H 3 acetylation in group B, C, and D were 1.12, 4.21and 4.38, respectively. The level of histone H 3 acetylation in group C and D was increased more compared with that in group A (P<0.05) and there was no difference between group B and group A ( P>0.05). The 2-Δ Δ Ct value of the expression of ING 1b mRNA in group B, C and D were1.33, 4.52and 4.62, respectively. The expression of ING1b mRNA in group C and D were more than that in group A ( P<0.05). Group B and group A had a similar level of ING 1b mRNA expression ( P>0.05). Conclusion:The histone acetylation is probably responsible for ING1b expression silencing in Colo205 human colon cell line. TSA at100 μ g/L can increase the level of acetylation and activate the gene transcription which is silenced by low level of acetylation and induce the expression of gene, inhibiting the growth of tumor cells.

     

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