摘要
目的探讨沉默缺氧诱导因子-1α(HIF-1α)对人舌鳞状细胞癌Tca8113细胞移植瘤血管生成拟态(VM)的影响及其可能的机制。方法 (1)沉默Tca8113细胞的HIF-1α,Real time RT-PCR和Western blot验证干扰效率;(2)建立裸鼠皮下移植瘤模型:分为阴性对照组和沉默HIF-1α的干扰组,观察沉默HIF-1α对肿瘤生长的影响并绘制生长曲线;(3)瘤体HE染色及免疫组化CD31染色观察血管形态、微血管密度(MVD)和VM的变化,免疫组化染色观察沉默HIF-1α对血管内皮生长因子(VEGF)、转化生长因子-β1(TGF-β1)蛋白表达的影响。结果 (1)移植瘤的生长曲线显示干扰组瘤体的生长速度和体积显著慢于和小于对照组(P<0.05);(2)与对照组相比,干扰组瘤体内的血管形态趋于正常且MVD减小(P<0.05),VM现象少见;(3)与对照组相比,干扰组瘤体组织的VEGF、TGF-β1表达显著减弱,差异具有统计学意义(P<0.05)。结论口腔鳞癌移植瘤中存在VM现象,VM的形成可能与HIF-1α对靶基因VEGF、TGF-β1的表达调控有关。
Objective To investigate the role of HIF-1α in vasculogenic mimicry(VM) in Tca8113 tongue carcinoma murine xenograft model.Methods (1)Constructing HIF-1α shRNA to silence human HIF-1α in Tca8113 cell,Western blot assay and Real time reverse transcriptase polymerase chain reaction(Real time RT-PCR)were used to validate the inhibitory rate.(2)We injected selected stable HIF-1α silenced and negative control Tca8113 cell into two different groups respectively:interference group and negative control group to establish murine xenograft model,observe growth state and drawing growth curve.(3)HE staining and CD31 immunohistochemical staining were used to observe the morphological changes of blood vessel,microvessel density(MVD)and the changes of VM,and the expression of HIF-1α downstream target genes VEGF,TGF-β1 were detected by immunohistochemical staining.Results (1)Growth curve showed the growth speed and tumor volume of interference group were significant slower and smaller than control group(P<0.05).(2)Compared with control group,the morphology of blood vessel in interference group was verging to normal,MVD and VM reduced significantly(P<0.05).(3)The expression of VEGF,TGF-β1 of the interference group was remarkable weaker than the control group.Conclusions VM exists in the xenograft model of oral squamous cell cancer.HIF-1α and its target genes VEGF,TGF-β1 involve in the formation of VM.
出处
《中华临床医师杂志（电子版）》
CAS
2012年第19期5814-5820,共7页
Chinese Journal of Clinicians(Electronic Edition)
基金
内蒙古自治区科学基金(20080404MS1111)
关键词
口腔肿瘤
RNA干扰
缺氧诱导因子1
Α亚基
血管生成
血管生成拟态
Mouth neoplasms
RNA interference
Hypoxia-inducible factor-1,alpha subunit
Angiogenesis
Vasculogenic mimicry
