尾叶香茶菜二萜类化合物B对人宫颈癌细胞的生长抑制作用及其机制

目的:观察尾叶香茶菜二萜类化合物B(DB)对人宫颈癌细胞株Hela细胞的生长抑制作用并探讨其机制。方法:处于对数生长期的Hela细胞分为正常对照组,0.5、1.0、2.0、4.0、8.0、16.0、32.0mg.L-1DB实验组,MTT法观察不同剂量DB作用24和48h后Hela细胞增殖抑制率,相差显微镜观察DB作用后Hela细胞形态的改变,RT-PCR和Western blotting分别观察DB作用后Hela细胞P53、P21、CDK2mRNA和蛋白表达水平。结果:MTT检测,24h时2.0、4.0、8.0DB组Hela细胞增殖抑制率分别为18.18%、28.89%和32.84%,48h时2.0、4.0、8.0DB组Hela细胞增殖抑制率分别为26.64%、47.03%和51.90%,呈时间-剂量依赖性(P<0.05或P<0.01);形态学观察,DB作用后Hela细胞变圆、体积缩小,有部分细胞漂浮;2.0、4.0和8.0mg.L-1DB分别作用Hela细胞24和48h后,P53和P21mRNA和蛋白表达水平明显高于正常对照组(P<0.01),CDK2mRNA和蛋白表达水平明显低于正常对照组(P<0.01)。结论:DB可抑制Hela细胞生长,其机制可能与上调细胞周期相关基因p53、p21的表达,下调CDK2表达,从而影响细胞从G1期进入S期有关。

Inhibitory effect of diterpenoid B derived from Rabdosia excise on growth of human cervical cancer cells and its mechanism

Objective To observe the effect of diterpenoid B derived from Rabdosia excise (DB) on the cell line of cervical cancer (Hela) and its related mechanism.Methods Hela cells at logarithmic growth phase were divided into control group and 0.5, 1.0, 2.0, 4.0, 8.0, 16, 0 and 32.0 mg·L~(-1) DB experimental groups, MTT assay was used to detect the inhibitory rate of cell growth at 24 and 48 h, and the morphologic changes of cells were observed by phase contrast microscope.RT-PCR was exploited to observe the mRNA expressions of p53, p21 and CDK2 of Hela cells.Immunocytochemical staining was utilized to detect the protein expressions of P53, P21 and CDK2.Results MTT result showed that the inhibitory rate of cell growth at 24 h in 2.0, 4.0 and 8.0 mg · mL~(-1)DB groups were 18.18%, 28.89%, and 32.84%;the inhibitory rates of cell growth at 48 h in 2.0, 4.0, and 8.0μg· mL~(-1) DB groups were 26.64%, 47.03%, and 51.90%;DB inhibited Hela cell growth in a dose- and time- dependent manner (P<0.05 or P<0.01). Hela cells after treated with DB manifested retarded proliferation, and became round and smaller in size, with some cells floating;the RT-PCR and Western blotting results showed that after treated with DB (2.0, 4.0, 8.0μg· mL~(-1)) for 24 and 48 h, the mRNA and protein expressions of P53 and P21 in DB groups were significantly higher than those in control group (P<0.01), meanwhile the mRNA and protein levels of CDK2 were lower than those in control group (P< 0.01). Conclusion DB can inhibit the proliferation of Hela cells, its mechanism may be related to up-regulating the expressions of P53 and P21, down-regulating the expression of CDK2, and furthermore influencing the transition from G_1 to S phase.