人参皂甙Rg3联合三氧化二砷对人肝癌细胞增殖性的影响

The effect of ginsenoside Rg3 combined with arsennic trioxide on the proliferation of human hepatocellular carcinoma

目的探讨人参皂甙Rg3联合三氧化二砷对肝癌裸鼠移植瘤模型的治疗作用,观察其对移植瘤肝癌细胞增殖性的影响。方法采用人肝癌细胞株Bel-7402细胞,种植于32只雄性裸鼠皮下,随机分成联合用药组、Rg3组(5mg/kg、10mg/kg、20mg/kg)、三氧化二砷组及对照组。实验自肿瘤接种第3天开始,人参皂甙Rg3(3个剂量),隔日灌胃,共20次;三氧化二砷40μg.kg-1.d-1,每日腹腔内注射,连续28d;对照组为生理盐水。停药1周后,脱颈处死,完整取出瘤块,检测肿瘤的重量,计算抑瘤率。采用免疫组织化学法检测增殖细胞核抗原(PCNA)在裸鼠肝癌移植瘤组织中的表达情况。结果三氧化二砷组抑瘤率(55.25%)高于Rg3组(5mg/kg亚组为32.24%、10mg/kg亚组为45.10%,20mg/kg亚组为50.00%),但随着Rg3浓度的升高,二者之间的差异明显减小;联合用药组中,5mg/kg亚组平均瘤重(0.58±0.15)g,抑瘤率61.84%;10mg/kg亚组平均瘤重(0.54±0.11)g,抑瘤率64.47%;20mg/kg亚组平均瘤重(0.49±0.12)g,抑瘤率67.76%,其抑瘤作用高于三氧化二砷组,差异有统计学意义(P<0.05)。三氧化二砷组抑制肝癌细胞增殖作用(PI值:38.3±5.3)强于Rg3组(PI值:5mg/kg亚组58.1±2.4;10mg/kg亚组51.8±5.3;20mg/kg亚组41.9±6.5),联合用药组(PI值:5mg/kg亚组38.6±8.4;10mg/kg亚组30.9±3.2;20mg/kg亚组23.3±3.3)作用最强(P<0.05),可以明显抑制移植瘤肝癌细胞增殖性。结论三氧化二砷对裸鼠肝癌移植瘤生长的抑制作用可能是通过抑制PCNA蛋白的表达。人参皂甙Rg3与三氧化二砷联合用药对移植瘤肝癌细胞增殖性的抑制有协同作用。

Objective To observe the therapeutic effect of ginsenoside Rg3 combined with arsennic trioxide（As2O3）on transplanted hepatic carcinoma in nude mice and to investigate its effect on the proliferation of hepatoma cells of transplanted carcinoma.Methods 32 male nude mice were randomly divided into 8 groups,and transplanted hepatic carcinoma models were set up by subcutaneously implanting human hepatic carcinoma cell strain（Bel-7402）.The mice in treatment groups were treated by Rg3（5 mg/kg,10 mg/kg,20 mg/kg）combined with As2O3,Rg3（5 mg/kg,10 mg/kg,20 mg/kg）alone,As2O3（40 μg·kg-1·d-1）alone and saline（0.5 ml,qd）respectively.3 days after the nude mice were inoculated,Rg3 was given by gastrogavage once every 2 days for 20 times,As2O3 was injected introperitoneally once a day for 28 days.A week after treatment being stopped,the mice were killed and the size of solid tumors was measured,tumor inhibition rate was calculated and intratumor PCNA was detected by immunohistochemical staining.Results The tumor inhibition rate（55.25%）in As2O3 group was higher than that（32.24%,45.10%,50.00% respectively）in Rg3 subgroups,but with the increase of dosage,the difference was not obvious.Among combined drug groups,in 5 mg/kg subgroup tumor mean weight was（0.58±0.15）g and tumor inhibition rate was 61.84%;in 10 mg/kg subgroup mean tumor weight was（0.54±0.11）g and tumor inhibition rate was 64.47%;in 20 mg/kg subgroup mean tumor weight was（0.49±0.12）g and inhibition tumor rate was 67.76%,that was significantly higher than that of As2O3 group（P〈0.05）.The inhibiting effect on the proliferation of hepatoma cells in As2O3 group（PI：38.3±5.3）was superior to that of Rg3 group（PI：58.1±2.4,51.8±5.3,41.9±6.5 respectively）.The effects in combined drug group（PI：38.6±8.4,30.9±3.2,23.3±3.3 respectively）were greatest,which could obviously inhibit the proliferation of hepatoma cells of transplanted tumor.Conclusion As2O3 can inhibit the growth of transplanted human hepatic carcinoma in nude mice mainly through inhibiting the expression of PCNA.Rg3 combined with As2O3 has synergistic effect in inhibiting the proliferation of hepatoma cells of transplanted tumor.