过表达TIPE2对肝癌生长和转移的影响  

Effect of Overexpression of TIPE2 on Growth and Metastasis of Liver Cancer

作　　者：孔丹丹 龚莲 谢宇锋[1] 陶敏[2] KONG Dandan;GONG Lian;XIE Yufeng(Department of Oncology,The First Affiliated Hospital of Soochow University,Jiangsu 215006,China)

机构地区：[1]苏州大学附属第一医院肿瘤科实验室,215006 [2]苏州大学附属独墅湖医院,215006

出　　处：《医学研究杂志》2023年第6期29-35,共7页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81772645)。

摘　　要：目的探讨肿瘤坏死因子诱导蛋白8样蛋白2(tumor necrosis factor-alpha-induced protein 8-like 2,TIPE2)对肝癌生长和转移的作用及调控机制。方法采用实时荧光定量PCR(real-time quantitative polymerase chain reaction,RT-qPCR)法检测TIPE2在肝癌细胞系和正常肝细胞(L02)中的表达;将本实验室构建的空对照腺病毒(AdV)及AdVTIPE2分别感染肝癌细胞,并检测其转染效率。用CCK-8、划痕愈合实验及Transwell侵袭实验检测肝癌细胞的增殖、迁移和侵袭能力。与人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVEC)共培养后,用小管形成实验检测细胞促血管形成能力,并观察HUVEC细胞的迁移、侵袭能力的变化。酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测细胞培养上清中血管内皮生长因子(vascular endothelial growth factor,VEGF)含量;Western blot法检测VEGF、缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP9)蛋白水平。结果TIPE2在肝癌细胞中表达降低;过表达TIPE2能显著抑制肝癌细胞的增殖、迁移和侵袭能力,以及内皮细胞的血管形成和迁移、侵袭能力。此外,TIPE2过表达能抑制VEGF的蛋白分泌,并且还可下调VEGF、HIF-1α和MMP9的蛋白表达水平。结论过表达TIPE2可抑制肝癌细胞的生长和转移,其机制可能与HIF-1α/VEGF信号通路失活及MMP9的表达下调,从而抑制新生血管形成有关。Objective To investigate the role and regulatory mechanism of tumor necrosis factor-inducible protein 8-like protein 2(TIPE2)on the growth and metastasis of liver cancer.Methods The expression of TIPE2 in liver cancer cell lines and normal hepatocytes(L02)was detected by Real-time quantitative PCR(RT-qPCR).The empty control adenovirus(AdV)and AdVTIPE2 constructed in our laboratory were respectively infected with hepatoma cells,and their transfection efficiency was tested.The proliferation,migration and invasion abilities of hepatoma cells were detected by CCK-8,scratch healing assay and Transwell invasion assay.After co-culture with human umbilical vein endothelial cells(HUVEC),the tubule formation assay was used to detect the ability of cells to promote angiogenesis.We observed the changes in the migration and invasion abilities of HUVECs.The content of vascular endothelial growth factor(VEGF)in the cell culture supernatant was detected by enzyme-linked immunosorbent assay(ELISA);Western blot was used to detect the protein levels of VEGF,hypoxia-inducible factor-1α(HIF-1α)and matrix metalloproteinase-9(MMP9).Results TIPE2 was lowly expressed in hepatoma cells;overexpression of TIPE2 could significantly reduce the proliferation,migration and invasion abilities of hepatoma cells,as well as the angiogenesis,migration,and invasion abilities of endothelial cells.In addition,overexpression of TIPE2 could inhibit the protein secretion of VEGF and down-regulate the protein expression levels of VEGF,HIF-1αand MMP9.Conclusion Overexpression of TIPE2 can inhibit the growth and metastasis of liver cancer cells,and the mechanism may be related to the inactivation of HIF-1α/VEGF signaling pathway and the down-regulation of MMP9 expression,thereby inhibiting the formation of new blood vessels.

关 键 词：TIPE2 生长和转移 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]