低氧条件下人参皂甙Rg3诱导人喉鳞癌细胞凋亡及对HIF-1α和VEGF表达的影响

Effect of Gensenoside Rg3 on Apoptosis and Expression of HIF-1α and VEGF in Hep2 Human Laryngeal Cancer Cell Line under Anoxic Conditions

目的研究人参皂甙Rg3对低氧条件下人喉鳞癌细胞(Hep-2)生长抑制作用以及可能的机制。方法通过体外低氧培养Hep-2人喉鳞癌细胞株,同时设立阴性对照组,阳性对照组(顺铂);采用TUNEL法(终末脱氧核苷酸末端转移酶介导缺口末端标记)检测细胞凋亡情况,计数凋亡细胞,计算凋亡率;流式细胞术测定细胞周期及细胞凋亡情况;应用免疫细胞化学技术和流式细胞技术检测Hep-2细胞在人参皂甙Rg3作用下HIF-1α和VEGF蛋白表达的变化;应用RT-PCR技术检测HIF-1α和VEGF mRNA表达的变化。结果TUNEL及流式细胞术检测Rg3组的凋亡率明显高于阴性对照组(P<0.05);在低氧条件下,Rg3组和顺铂对照组的Hep-2细胞中的HIF-1α和VEGF蛋白表达低于阴性对照组(P<0.05);Rg3组的HIF-1α和VEGF mRNA水平明显低于阴性对照组(P<0.05),顺铂组的HIF-1α和VEGF mRNA水平无明显变化。结论低氧条件下,人参皂甙Rg3诱导细胞凋亡,抑制了Hep-2细胞中的HIF-1α和VEGF蛋白和mRNA的表达,这可能是Rg3抗肿瘤作用的机制之一。

Objective To study the mechanism and effect of gensenoside Rg3 on Hep2 Cell Line during the normoxia and hypoxia. Methods Anoxic cultured Hep2 human laryngeal cancer cell line, mean-while the normal control group and positive control group （DDP）were set. The cell apoptotic was detected by TUNEL method. The cell cycle and cell apoptosis analysis was detected by FCM. Then the expression of HIF-1α and VEGF protein was detected by immunohistochemistry and flow cytometry; the expression of HIF-1α and VEGF mRNA was detected by transcription-polymerase chain reaction（RT-PCR）. Results The apoptosis rate obviously increase by TUNEL and FCM（P〈0. 05）. Compared with the Hep2 cells without any induction, the expression of HIF-1 a and VEGF protein in Rg3 groups and DDP groups obviously depressed（P〈0. 1）5）;the expression of HIF-1α and VEGF mRNA in Rg3 groups obviously depressed（P〈0.05）;but had no effect on the level of HIF-1α and VEGF mRNA in DDP groups. Conclusion Rg3 could inhibit the high protein and mRNA expression of HIF-1α and VEGF and it could induce cell apoptosis under anoxic conditions,which may be one of its anticancer mechanisms.