吉西他滨对胰腺癌PANC-1细胞中组蛋白去乙酰化酶及TMS1/ASC基因表达的影响

Effects of gemcitabine on the expression of TMS1 /ASC gene and histone deacetylase in pancreatic carcinoma cell line PANC-1

目的检测吉西他滨(GEM)对胰腺癌细胞株PANC-1中甲基化诱导静止基因(TMS1/ASC)及组蛋白去乙酰化酶(HDAC)表达的影响,并探讨组蛋白去乙酰化与TMS1/ASC之间的关系。方法 4.27μg/ml GEM作用于PANC-1细胞,运用透射电镜观察GEM对胰腺癌细胞PANC-1形态的影响。蛋白印迹法(Western blotting)检测HDAC的表达,用重亚硫酸盐测序PCR(BSP)和结合重亚硫酸盐的限制性内切酶法(COBRA)检测TMS1/ASC启动子区域甲基化状态。结果 TMS1/ASC在胰腺癌细胞株PANC-1中发生甲基化,经5-杂氮-2'-脱氧胞苷(5-aza-dC)处理后,PANC-l细胞TMS1/ASC重新表达,GEM诱导胰腺癌细胞株PANC-1中TMS1/ASC未发生甲基化。HDAC在GEM诱导胰腺癌细胞株PANC-1中表达水平低于正常对照组。结论 GEM能抑制胰腺癌PANC-1细胞增殖并促进凋亡,随着作用时间的延长,其药物敏感性减低。GEM可能通过促进组蛋白发生乙酰化和抑制TMS1/ASC启动子甲基化来治疗胰腺癌。

Objective To investigate the expression of TMS1/ASC gene and histone deacetylase（HDAC） in pancreatic carcinoma cell line PANC-1 treated with gemcitabine （GEM）, and to investigate the relationship between histone acetylation and the expression of TMS1/ASC. Methods The pancreatic carcinoma cell line PANC-1 was incubated in DMEM, and the cells were treated with 4. 27μg/ml GEM. The morphological changes of the neutrophils were evaluated by electron microscopy. Western biotting analysis was performed to detect the expression of HDAC. Bisulfite sequencing PCR （BSP） and combined bisulfite restriction analysis （COBRA）was used to test promoter methylation status. Results TMS1/ASC promoter metbylation was not significantly changed after GEM treatment. The expression of HDAC was downregulated in PANC-1 treated with GEM. Conclusions GEM can inhibit the proliferation of PANC-1 cells and induce their apoptosis. The apoptosis of PANC-1 cells induced by GEM might be dependent on promoting histone acetylation and independent of methylation status of TMS1/ASC gene.