奈达铂对宫颈癌Hela及Siha细胞的作用

Effect of nedaplatin on cervical cancer HeLa cells and SiHa cells

目的:比较奈达铂(NDP)体外对人宫颈腺癌细胞Hela细胞及鳞癌细胞Siha的抑制作用并探讨其作用机制。方法:以2.5～40μg/ml NDP分别作用于Hela及Siha细胞24 h,48 h,72 h,MTT法检测抑制率,计算半数抑制浓度(IC50);利用流式细胞术(FCM)观察细胞凋亡率及周期变化;半定量PCR法分析基因半胱氨酸天门冬氨酸蛋白酶3(Caspase 3,Caspase 9)及基质金属蛋白酶2(MMP-2)的表达变化。结果:NDP对宫颈癌Hela细胞及Siha细胞均有抑制作用,呈时间-剂量依赖性,NDP对Hela细胞的抑制率大于Siha细胞;FCM显示,随时间、浓度增加,Hela及Siha细胞凋亡率增加,相同浓度和作用时间下,Hela细胞的凋亡率大于Siha细胞(P<0.05),S期细胞的比例增加,G0/G1期比例减少,G2/M期比例变化不明显;与对照组相比,Caspase 3,Caspase 9表达增加,MMP-2表达减少。结论:NDP可抑制宫颈腺癌细胞Hela及鳞癌细胞Si-ha的增殖,NDP体外对Hela细胞的抑制作用更强。NDP的作用机制与诱导细胞凋亡、细胞周期阻滞、上调Caspase3和Caspase9的表达有关;下调MMP-2的表达可能有利于降低宫颈癌细胞的侵袭力。

Objective： To compare the inhibiting effect of nedaplatin in vitro on human cervical adenocarcinoma HeLa cells and cervical squamous carcinoma SiHa cells and explore the mechanism.Methods： HeLa cells and SiHa cells were treated with nedaplatin（2.5-40 μg/ml） for 24,48 and 72 hours,respectively,MTT method was used to detect the inhibition rate,and then the median inhibitory concentration（IC50） was calculated;flow cytometry was used to observe the apoptotic rate and change of cycles;semiquantitative PCR was used to analyze the changes of expressions of Caspase 3,Caspase 9 and matrix metal proteinase-2（MMP-2）.Results： Nedaplatin inhibited cervical cancer HeLa cells and SiHa cells,showing a time-dose-dependent manner,the inhibition rate of nedaplatin on HeLa cells was higher than that on SiHa cells;the results of flow cytometry showed that the apoptotic rates of HeLa cells and SiHa cells increased with the increase of concentration,the apoptotic rate of HeLa cells was statistically significantly higher than that of SiHa cells under the same concentration and action time（P 0.05）,the proportion of cells at S phase increased,and the proportion of cells at G0 / G1 phase decreased,the change of proportion of cells at G2 / M phase was not significant;compared with control group,the expression levels of Caspase 3 and Caspase 9 increased,while the expression level of MMP-2 decreased.Conclusion： Nedaplatin can inhibit the proliferation of cervical cancer HeLa cells and SiHa cells,application of nedaplatin in vitro has more stronger inhibiting effect on HeLa cells.The mechanism is correlated with inducing apoptosis,cell cycle arrest,up-regulation of Caspase3 and Caspase9;down-regulation of MMP-2 may be helpful to reduce the invasive ability of cervical cancer cells.