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目的:观察组蛋白乙酰基转移酶(histone deacetylase,HDAC)抑制剂曲古抑菌素A(trichostatin A,TSA)对子宫内膜癌Ishikawa细胞凋亡的影响,并研究其与Krupell样因子4(Krupell-like factor 4,KLF4)的关系。方法:0、50、100、200、300、500ng/ml TSA作用于Ishikawa细胞24 h,或100 ng/ml TSA作用于Ishikawa细胞0、4、8、12、24、48 h,流式细胞术检测Ishikawa细胞凋亡情况,qRT-PCR检测Ishikawa细胞中KLF4 mRNA的表达情况;将KLF4真核表达载体pcDNA3-KLF4转染Ishikawa细胞,流式细胞术检测Ishikawa细胞凋亡情况。结果:100 ng/ml TSA作用于Ishikawa细胞24 h后,Ishikawa细胞的凋亡率显著高于对照组[(30.6±4.5)%vs(7.53±0.93)%,P<0.05];不同质量浓度TSA处理Ishikawa细胞24 h后或100 ng/ml TSA作用Ishikawa细胞不同时间后,KLF4 mRNA表达水平以剂量依赖和时间依赖方式明显增高(P<0.05);pcDNA3-KLF4转染后Ishikawa细胞凋亡率显著增加[(27.3±2.7)%vs(4.53±1.75)%,P<0.05]。结论:TSA能通过诱导子宫内膜癌Ishikawa细胞中KLF4的表达,促进Ishikawa细胞发生凋亡。
OBJECTIVE: To observe the effect of histone deacetylase (HDAC) inhibitor trichostatin A (TSA) on the apoptosis of endometrial carcinoma cell line Ishikawa and to study its relationship with Krupell-like factor 4 ( Krupell-like factor 4, KLF4). Methods: Ishikawa cells were treated with 0, 50, 100, 200, 300 and 500 ng / ml TSA for 24 h or 100 ng / ml TSA for 0, 4, 8, 12, 24 and 48 h. Flow cytometry The apoptosis of Ishikawa cells was detected by qRT-PCR. The expression of KLF4 mRNA in Ishikawa cells was detected by qRT-PCR. The Ishikawa cells were transfected with the pcDNA3-KLF4 KLF4 eukaryotic expression vector. Flow cytometry was used to detect the apoptosis of Ishikawa cells. Results: The apoptosis rate of Ishikawa cells was significantly higher in Ishikawa cells treated with 100 ng / ml TSA [(30.6 ± 4.5)% vs (7.53 ± 0.93)%, P <0.05] After treatment with Ishikawa cells for 24 h or 100 ng / ml TSA treatment, the expression of KLF4 mRNA in Ishikawa cells was significantly increased in a dose-and time-dependent manner (P <0.05). The apoptosis rate of Ishikawa cells was significantly increased after transfection with pcDNA3-KLF4 Increased by (27.3 ± 2.7)% vs (4.53 ± 1.75)%, P <0.05. Conclusion: TSA can promote the apoptosis of Ishikawa cells by inducing the expression of KLF4 in Ishikawa cells of endometrial carcinoma.