青天葵总黄酮对人鼻咽癌CNE-2荷瘤裸鼠皮下肿瘤的抑制作用

The tnhibitory effect of Total Flavonoids from Nervilia Fordii on subcutaneous tumors of human nasopharyngeal carcinoma CNE-2 in tumor-bearing nude mice

目的:研究青天葵总黄酮(TFNF)对BALB/c裸鼠人鼻咽癌细胞皮下种植模型中TGF-β1/Erk/MAPK通路的调控作用。方法:BALB/c裸鼠皮下接种CNE-2细胞(1.0×107细胞/ml)建立鼻咽癌荷瘤模型,苏木精-伊红染色(HE)行病理判断,免疫组化法(IHC)观察细胞周期调节蛋白-67(Ki67)、增殖细胞核抗原(PCNA)的表达水平。免疫印迹法检测转化生长因子-β1(TGF-β1)、细胞外调节蛋白激酶(Erk)、p-Erk、丝裂原活化蛋白激酶(MAPK)、p-MAPK的蛋白表达水平。试剂盒法评估肾脏、肝脏、脾脏中超氧化物歧化酶(SOD)及丙二醛(MDA)水平。结果:与模型组比较,高剂量TFNF显著抑制荷瘤小鼠肿瘤体积与质量,显著降低Ki67(55.19%)、PCNA(46.65%)表达水平,提高肾脏、肝脏、脾脏中SOD水平至3.88、2.98、3.71U/mgprot,降低肾脏、肝脏、脾脏的MDA水平至2.19、1.05、2.85nmol/mgprot。TFNF可降低肿瘤组织中TGF-β1(47.36%)、p-Erk/Erk(81.13%)、p-MAPK/MAPK(19.80%)的蛋白表达水平。结论:TFNF可调控TGF-β1/Erk/MAPK通路抑制肿瘤生长,并减轻荷瘤裸鼠主要脏器的氧化损伤发生。

Objective:The regulatory effect of Total Flavonoids from Nervilia fordii(TFNF)on the TGF-β1/Erk/MAPK pathway in the subcutaneous transplantation model of human nasopharyngeal cancer cells in BALB/c nude mice was investigated.Methods:BALB/c nude mice were inoculated with CNE-2 cells(1.0×107cells/ml)to establish a tumor-bearing model of nasopharyngeal carcinoma,the pathological determination was performed by hematoxylin-eosin(HE),and the expression of cell cycle regulatory protein-67(Ki67)and proliferation nuclear antigen(PCNA)were observed by immunohistochemistry(IHC).Western blotting detected the protein expression of transforming growth factor-β1(TGF-β1),extracellular regulated protein kinase(Erk),p-Erk,mitogen-activated protein kinase(MAPK),and p-MAPK.Superoxide dismutase(SOD)and malondialdehyde(MDA)levels in the kidneys,liver,and spleen were assessed by the kit.Results:Compared with the model group,high-dose TFNF significantly inhibited tumor volume and weight,significantly reduced the expression levels of Ki67(55.19%)and PCNA(46.65%),increased SOD levels in kidneys,liver,and spleen(to 3.88,2.98,3.71 U/mg prot),and decreased MDA levels in kidneys,liver,and spleen(to 2.19,1.05,2.85 nmol/mg prot).In addition,TFNF can also reduce the protein expression levels of TGF-β1(47.36%),p-Erk/Erk(81.13%),and p-MAPK/MAPK(19.80%)in tumor tissues.Conclusion:The results showed TFNF could regulate the TGF-β1/Erk/MAPK pathway,inhibit tumor growth,and reduce the occurrence of oxidative damage to the main organs of tumor-bearing nude mice.