5-氮胞苷对HSG细胞的诱导凋亡作用

目的体外研究5-氮胞苷（5-azacytidine,5-azaC）对起源于人颌下腺闰管的HSG（human salivary gland,HSG）细胞的诱导凋亡作用,为应用5azaC对涎腺肿瘤进行治疗奠定理论和实验基础。方法用不同浓度的5-azaC处理HSG细胞,观察5-azaC在非毒性浓度下,能否诱导HSG细胞发生凋亡。结果加入5-azaC后48h,光镜下可见到少数细胞体积缩小,胞浆向外突起,类似出芽及发泡状;琼脂糖凝胶电泳结果显示经5-azaC作用72h的基因组DNA呈现出＂梯状＂图像,基因组DNA被内切酶切成了180bp～200bp及其整倍数的片断;流式细胞仪检测结果显示细胞在G1期前面出现亚二倍体峰;透射电镜结果显示5-azaC处理组细胞可见核内凋亡小体;免疫组化结果显示5-azaC可以抑制P53、Bcl-2和FAP-1的表达;Western blot结果显示5-azaC处理后可使FAP-1的表达降低。结论5-azaC在体外能诱导HSG细胞凋亡。

The effect of apoptosis induced by 5-azaC on HSG cells in vitro

Objective To investigate the effect of apoptosis induced by 5-azaC on HSG cells in order to establish basis for the treatment of salivary gland tumor.Methods HSG cells were treated with different concentration of 5-azaC. Morphology,flowcytotometry,DNA ladder,TME and immunohistochemistry were observed on HSG cells to detect if HSG cells underwent apoptosis.Results Growth of HSG cells was remarkably inhibited by 5-azaC,and when treated with 5uM 5-azaC,HSG cells began to undergo apoptosis detected by flowcytotometry and TEM. HSG cells showed apoptosis peaks when measured by flowcytometry and TEM showed margination of nuclear chromatin and shrinkage of cell body with cytoplasmic vacuolation.Immunohistochemistry results showed that 5-azaC could inhibit the expression of P53,Bcl-2 and FAP.Western blot results showed that 5azaC could inhibit the expression of FAP-1.Conclusion 5-azaC can induce apoptosis of HSG cells in vitro.