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目的探讨凋亡抑制蛋白生存素在紫杉醇诱导肝癌细胞株SMMC-7721凋亡中的作用。方法用紫杉醇浓度为1、10、100、1000、10 000ng/ml(分别为A、B、C、D、E组)诱导肝癌细胞株SMMC-7721 24h后,采用MTT法检测细胞抑制率,台盼蓝染色观察细胞凋亡情况;另设空白对照(F)组。以紫杉醇0、0.375、3.75、37.5、375、3750ng/ml诱导肝癌细胞株SMMC-7721 24h,采用RT-PCR和Western blot检测生存素的mRNA和蛋白表达。结果 A、B、C、D、E组的细胞抑制率分别为10.2%、37.1%、65.5%、86.3%、92.7%。B、C、D、E组细胞凋亡率均高于F组[(18.2±3.0)%、(53.5±4.7)%、(73.6±3.4)%、(84.2±5.6)%vs.(3.3±1.6)%](P<0.05)。当诱导浓度≤3.75ng/ml时,生存素mRNA及蛋白水平随浓度增加而升高;而当诱导浓度≥37.5ng/ml时,生存素mRNA及蛋白水平随浓度增加而下降(P<0.05)。结论生存素表达的下调可能促进紫杉醇诱导肝癌细胞株SMMC-7721凋亡的作用。
Objective To investigate the role of apoptosis inhibitor survivin in paclitaxel-induced hepatoma cell line SMMC-7721 apoptosis. Methods The hepatocellular carcinoma cell line SMMC-7721 was induced with paclitaxel at 1, 10, 100, 1000 and 10 000 ng / ml (groups A, B, C, D and E, respectively) Trypan blue staining observed cell apoptosis; another set of blank control (F) group. The hepatoma cell line SMMC-7721 was induced with paclitaxel at 0,0.375,3.75,37.5,375,3750ng / ml for 24 hours. The mRNA and protein expression of survivin were detected by RT-PCR and Western blot. Results The cell inhibitory rates in groups A, B, C, D and E were 10.2%, 37.1%, 65.5%, 86.3% and 92.7%, respectively. The apoptotic rates in group B, C, D and E were significantly higher than those in group F [(18.2 ± 3.0)%, (53.5 ± 4.7)%, (73.6 ± 3.4)%, (84.2 ± 5.6)% vs 1.6%] (P <0.05). Survivin mRNA and protein levels increased with increasing concentration when induced concentration was less than 3.75ng / ml, while survivin mRNA and protein levels decreased with increasing concentration when the induced concentration was greater than or equal to 37.5ng / ml (P <0.05) . Conclusions The down-regulation of survivin expression may promote paclitaxel-induced apoptosis in hepatocellular carcinoma cell line SMMC-7721.