水蛭素对人鼻咽癌细胞增殖的影响及机制

The proliferation inhibition effect with hirudin on human nasopharyngeal cancer CNE2 cells

目的探讨水蛭素对人鼻咽癌细胞(CNE2)增殖的影响及机制。方法取对数生长期的CNE2细胞随机分为对照组与水蛭素组。对照组常规培养,水蛭素组加入不同浓度的水蛭素进行培养。倒置显微镜下观察细胞形态学变化;MTT法测算CNE2细胞生长抑制率,计算不同时间点半抑制浓度(IC_(50));实时荧光定量PCR检测CNE2细胞内Bax、p21 mRNA表达。结果不同浓度不同时间点水蛭素组生长抑制率均高于对照组,水蛭素组各浓度各时间点生长抑制率比较差异均有统计学意义(P均<0.05)。药物作用24、48、72 h的IC_(50)分别为(8.28±2.1)、(5.11±0.7)、(4.83±0.5)ATU/mL,48、72 h时间点IC_(50)比24 h高(P均<0.05)。与对照组比较,5、8 ATU/mL水蛭素组Bax、p21 mRNA相对表达量高(P均<0.05),8 ATU/mL水蛭素组最高(P均<0.05)。结论水蛭素可明显抑制CNE2细胞增殖,该作用可能与水蛭素上调Bax、p21 mRNA表达有关。

Objective To investigate the proliferation inhibition effect of hirudin on human nasopharyngeal cancer CNE2 cells.Methods The human nasopharyngeal carcinoma CNE2 were cultured with common cell culture technique,and assigned to control group(culture medium+cells)and observation group(culture medium+cells+hirudin).Morphological changes of cells were observed under inverted microscope.Inhibitory effect of cell proliferation was measured by MTT assay,and calculation of inhibition concentration at different time(IC 50).Quantitative real-time polymerase chain reaction was used to demonstrate expression levels of involved mRNA such as p21,Bax.Results Compared with the control group,CNE2 cells in the observation group were cultured at 5 ATU/mL and 8 ATU/mL for 24 h and 48h,respectively,and then increased with the time and the dosing concentration,the cells adhered to the wall decreased,debris increased,and apoptosis was obvious.Hirudin could inhibit CNE2 cells proliferation,with the increasing of concentration and time of hirudin on CNE2 cells(all P<0.05).The IC 50 at 24 h,48 h and 72 h was(8.28±2.1),(5.11±0.7)and(4.83±0.5)ATU/mL respectively.The apoptosis rate of the group treated by hirudin raised in evidence(all P<0.05).The expression levels of p21 and Bax mRNA were significantly increased with hirudin(all P<0.05).Conclusion Hirudin significantly inhibits the proliferation of CNE2 cells,which may be related to the up-regulation of p21and Bax mRNA expression by hirudin.