摘要
目的探讨在缺氧微环境下Pokemon对肝细胞癌血管形成的影响。方法选取肝细胞癌细胞株HepG2和Huh7,采用短发夹RNA稳定转染并筛选稳定沉默Pokemon基因表达的肝细胞癌细胞株。通过RT-PCR验证沉默Pokemon之后血管形成相关基因的表达。在缺氧条件下通过Western blot法验证Pokemon基因沉默后缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)表达的影响。结果成功构建Pokemon干扰质粒。HepG2和Huh7细胞在Pokemon基因沉默后,血管形成相关基因促血管生成素2(angiopoietin-2,ANG2)、血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)以及血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2)的mRNA表达均受到抑制(均P<0.05)。在缺氧条件下,HepG2和Huh7细胞在Pokemon基因沉默后HIF-1α蛋白表达均下调(均P<0.05)。将HIF-1α质粒瞬转至Hela细胞36 h后,通过Western blot检测发现HIF-1α的表达增加(P<0.05),而Pokemon的表达变化差异没有统计学意义(P>0.05)。结论在缺氧微环境下Pokemon上调HIF-1α表达促进肝细胞癌血管形成,促进肝细胞癌的发生及发展。
Objective To investigate the effect of Pokemon on the angiogenesis of hepatocellular carcinoma cells under hypoxia microenvironment.Methods Hepatocellular carcinoma HepG2 and Huh7 cell lines were stably transfected with short hairpin RNA to establish Pokemon-silenced hepatocellular carcinoma cell lines.The expressions of angiogenesis-related genes were confirmed by RT-PCR after Pokemon silencing.The effect of Pokemon silencing on hypoxia inducible factor-1α(HIF-1α)expression was tested by Western blot under hypoxia condition.Results A plasmid encoding a short interfering RNA(siRNA)targeting Pokemon was constructed.Silencing of Pokemon inhibited angiogenesis related mRNA expressions in HepG2 and Huh7 cell lines,including mRNA of angiopoietin-2(ANG2),vascular endothelial growth factor A(VEGFA)and vascular endothelial growth factor receptor 2(VEGFR2;all P<0.05).Under hypoxia condition,the HIF-1αexpression was down-regulated in HepG2 and Huh7 cells after Pokemon silencing(both P<0.05).When a plasmid encoding siRNA targeting HIF-1αwas transitioned into HeLa cells 36 h later,the expression of HIF-1αprotein was increased(P<0.05),but the expression of Pokemon protein was unchanged(P>0.05).Conclusions The up-regulation of HIF-1αexpression by Pokemon in hypoxic microenvironment promotes the angiogenesis of hepatocellular carcinoma cells and leads to the occurrence and development of hepatocellular carcinoma.
作者
张玉琴
陈章兴
戴益琛
朱小三
詹晓娟
傅育卡
吴婧
Zhang Yuqin;Chen Zhangxing;Dai Yichen;Zhu Xiaosan;Zhan Xiaojuan;Fu Yuka;Wu Jing(Department of Gastroenterology,Chenggong Hospital of Xiamen University,Xiamen 361003,China)
出处
《实用肿瘤杂志》
CAS
2022年第5期444-449,共6页
Journal of Practical Oncology
基金
厦门市医疗卫生科技计划项目(3502Z20184067)。
