Aglaroxin C对肝癌细胞体内外生长的抑制作用

Inhibiting effect of Aglaroxin C on growth of hepatocellular carcinoma in vitro and in vivo

目的探讨Aglaroxin C对人肝癌细胞HepG2体外增殖的影响,并初步探讨其对肝癌移植瘤小鼠的抗肿瘤作用。方法体外培养人肝癌细胞HepG2,分别加入1、10、100 nmol·L^(-1)及1、10、100μmol·L^(-1)Aglaroxin C作用24、48、72 h,采用细胞计数试剂盒(CCK-8)检测肝癌细胞HepG2的增殖抑制率,并计算Aglaroxin C的半数抑制浓度(IC50)。取25只Balb/c小鼠随机分为尾静脉对照组、尾静脉0.10 mg Aglaroxin C组、瘤内对照组、瘤内0.05 mg Aglaroxin C组、瘤内0.10 mg Aglaroxin C组,每组5只。各组小鼠左腋皮下接种鼠肝癌细胞H22建立肝癌移植瘤模型,模型建立成功后,尾静脉对照组、尾静脉0.10 mg Aglaroxin C组小鼠分别通过尾静脉注射生理盐水0.2 mL、0.50 g·L^(-1)Aglaroxin C 0.2 mL;瘤内对照组、瘤内0.05 mg Aglaroxin C组、瘤内0.10 mg Aglaroxin C组小鼠分别瘤内多点注射生理盐水0.2 mL、0.25 g·L^(-1)Aglaroxin C 0.2 mL、0.50 g·L^(-1)Aglaroxin C 0.2 mL;隔日1次,共4次。给药期间监测小鼠肿瘤体积变化;末次给药后24 h,剖取各组小鼠肿瘤组织,称质量并计算肿瘤抑制率;取肿瘤组织行苏木精-伊红染色,观察各组小鼠肿瘤组织病理学变化。结果不同浓度Aglaroxin C干预人肝癌细胞HepG2 24、48、72 h后,同一时间点人肝癌细胞HepG2的增殖抑制率随Aglaroxin C浓度的增加而升高(P<0.05);Aglaroxin C干预人肝癌细胞HepG224、48、72 h的IC50分别为73、25、82 nmol·L^(-1)。给药干预2~8 d,尾静脉0.10 mg Aglaroxin C组小鼠肿瘤体积小于尾静脉对照组(P<0.05);瘤内0.05 mg Aglaroxin C组、瘤内0.10 mg Aglaroxin C组小鼠肿瘤体积均小于瘤内对照组(P<0.05);瘤内0.05 mg Aglaroxin C组小鼠肿瘤体积与尾静脉0.10 mg Aglaroxin C组比较差异无统计学意义(P>0.05),瘤内0.10 mg Aglaroxin C组小鼠肿瘤体积小于尾静脉0.10 mg Aglaroxin C组(P<0.05)。给药干预5~8 d,瘤内0.10 mg Aglaroxin C组小鼠肿瘤体积小于瘤内0.05 mg Aglaroxin C组(P<0.05)。尾静脉0.10 mg Aglaroxin C组小鼠肿瘤质量明显小于尾静脉对照组(P<0.05);瘤内0.05 mg Aglaroxin C组小鼠肿瘤质量与尾静脉0.10 mg Aglaroxin C组比较差异无统计学意义(P>0.05),瘤内0.10 mg Aglaroxin C组小鼠肿瘤质量小于尾静脉0.10 mg Aglaroxin C组和瘤内0.05 mg Aglaroxin C组(P<0.05)。瘤内0.05 mg Aglaroxin C组小鼠肿瘤质量和肿瘤抑瘤率与尾静脉0.10 mg Aglaroxin C组比较差异无统计学意义(P>0.05)。瘤内0.10 mg Aglaroxin C组小鼠肿瘤抑瘤率高于尾静脉0.10 mg Aglaroxin C组和瘤内0.05 mg Aglaroxin C组(P<0.05)。显微镜下可见,瘤内0.10 mg Aglaroxin C组小鼠肿瘤组织坏死程度最明显。结论 Aglaroxin C可以显著抑制人肝癌细胞HepG2的体外增殖;对肝癌移植瘤小鼠具有显著的抗肿瘤活性。

Objective To explore the inhibiting effect of Aglaroxin C on growth of hepatocellular carcinoma HepG2 in vitro and to evaluate its antitumor activity in tumor-bearing mice.Methods The human hepatocellular carcinoma cell line HepG2 were cultured in vitro and the cells were intervened by 1,10,100 nmol·L-1 and 1,10,100μmol·L-1 Aglaroxin C for 24,48,72 h,respectively.The proliferation inhibition rate of human hepatocellular carcinoma cell line HepG2 was determined by cell counting kit-8 assay,and 50%inhibiting concentration of the Aglaroxin C was calculated.Twenty-five Balb/c mice were selected and randomly divided into tail vein control group,0.10 mg Aglaroxin C tail vein group,intratumoral administration control group,0.05 mg Aglaroxin C intratumoral administration group and 0.10 mg Aglaroxin C intratumoral administration group,with five mice in each group.Transplanted tumor model of hepatocellular carcinoma was established by subcutaneous inoculation of hepatocellular carcinoma cells H22 in left axilla of mice in each group.Subsequently,the mice in the tail vein control group and 0.10 mg Aglaroxin C tail vein group were injected 0.2 mL physiological saline through tail vein and 0.2 mL Aglaroxin C(0.50 g·L-1),respectively.The mice in the intratumoral administration control group,0.05 mg Aglaroxin C intratumoral administration group and 0.10 mg Aglaroxin C intratumoral administration group were multipoint injected 0.2 mL physiological saline,0.2 mL Aglaroxin C(0.25 g·L-1)and 0.2 mL Aglaroxin C(0.50 g·L-1)into intratumor.Once every other day,four times in total.Changes of tumor volume in mice were measured during administration.Twenty-four hours after the last drug administration,tumor tissues of the mice in each group were also dissected out,the mass of tumor was measured,the tumor inhibition rate was calculated.Part of the tumor tissue was used to observe the pathological changes by hematoxylineosin staining.Results At 24,48,72 h after treating the human hepatocellular carcinoma cell line HepG2 by Aglaroxin C with different concentration,the proliferation inhibition rate of human hepatocellular carcinoma cell line HepG2 increased with the increasing of Aglaroxin C concentration(P<0.05),and the IC 50 of the Aglaroxin C on the human hepatocellular carcinoma cell line HepG2 cells was 73,25,82 nmol·L-1.At 2-8 days of administration,the tumor volume of 0.10 mg Aglaroxin C tail vein group was smaller than that of the tail vein control group(P<0.05);the tumor volume of 0.05 mg Aglaroxin C intratumoral administration group and 0.10 mg Aglaroxin C intratumoral administration group was smaller than that of the intratumoral administration control group(P<0.05).There was no significant difference in the tumor volume between the 0.05 mg Aglaroxin C intratumoral administration group and 0.10 mg Aglaroxin C tail vein group(P>0.05);the tumor volume of 0.10 mg Aglaroxin C intratumoral administration group was smaller than that of 0.10 mg Aglaroxin C tail vein group(P<0.05).At 5-8 days of administration,the tumor volume of 0.10 mg Aglaroxin C intratumoral administration group was smaller than that of 0.05 mg Aglaroxin C intratumoral administration group(P<0.05).The tumor mass of 0.10 mg Aglaroxin C tail vein group was smaller than that of the tail vein control group(P<0.05);the tumor mass of 0.05 mg and 0.10 mg Aglaroxin C intratumoral administration group was smaller than that of the intratumoral administration control group(P<0.05);there was no significant difference in the tumor mass between the 0.05 mg Aglaroxin C intratumoral administration group and 0.10 mg Aglaroxin C tail vein group(P>0.05);the tumor mass of 0.10 mg Aglaroxin C intratumoral administration group was smaller than that of 0.05 mg Aglaroxin C intratumoral administration group and 0.10 mg Aglaroxin C tail vein group(P<0.05).There was no significant difference in the tumor mass and inhibition rate between the 0.05 mg Aglaroxin C intratumoral administration group and 0.10 mg Aglaroxin C tail vein group(P>0.05).The tumor inhibition rate of 0.10 mg Aglaroxin C intratumoral administration group was higher than that of 0.10 mg Aglaroxin C tail vein group and 0.05 mg Aglaroxin C intratumoral administration group(P<0.05).Under the microscope,the degree of tumor necrosis of the mice in 0.10 mg Aglaroxin C intratumoral administration group was the most obviously.Conclusion Aglaroxin C treatment can significantly inhibit the proliferation of human hepatocellular carcinoma cell line HepG2 in vitro.It has the antineoplastic activity on the tumor-bearing mice.