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目的:探讨缺氧诱导因子-1α(HIF-1α)基因沉默对人食管鳞癌细胞体内血管生成拟态相关基因表达的影响。方法:将食管鳞癌Eca109细胞株分为空白对照组(未转染组)、阴性对照组(转染空质粒组)和实验组(稳定转染重组质粒pGCsi-shHIF-1α组),接种裸鼠,观测裸鼠皮下移植瘤生长情况;HE染色观察各组肿瘤的异型性及坏死情况;免疫组化和Western blot法检测HIF-1α与上皮细胞激酶(EPHA2)、血管上皮钙黏附素(VE-cadherin)、基质金属蛋白酶-2(MMP-2)在肿瘤组织中的蛋白表达;RT-PCR检测移植瘤组织中HIF-1α与EPHA2、VE-cadherin、MMP-2的mRNA表达。结果:实验组出瘤时间延迟,生长速度减慢,移植瘤体积及重量明显减少(P<0.05);HE染色示肿瘤异型性减少,坏死显著增多;免疫组化及Western blot发现实验组HIF-1α蛋白表达完全沉默,EPHA2、VE-cadherin表达明显减少(P<0.05);RT-PCR提示实验组中HIF-1α及EPHA2、VE-cadherin、MMP2的mRNA含量均有不同程度减少,其中EPHA2、VE-cadherin减少明显(P<0.05)。结论:HIF-1α沉默能显著抑制食管鳞癌裸鼠移植瘤的生长,并能有效抑制移植瘤血管生成拟态相关基因EPHA2和VE-cadherin的表达,HIF-1α有可能通过以上2个位点参与食管鳞癌血管生成拟态的调控。
Objective: To investigate the effect of hypoxia inducible factor-1α (HIF-1α) gene silencing on the expression of vasculogenic mimicry related gene in human esophageal squamous cell carcinoma. Methods: Eca109 cells were divided into blank control group (untransfected group), negative control group (transfected empty plasmid group) and experimental group (stably transfected with recombinant plasmid pGCsi-shHIF-1α group) The growth of subcutaneous xenograft in nude mice was observed. The heterogeneity and necrosis of tumor in each group were observed by HE staining. The expressions of HIF-1α, EPHA2, VE The protein expression of MMP-2 and MMP-2 in tumor tissue was detected by RT-PCR. The mRNA expressions of HIF-1α, EPHA2, VE-cadherin and MMP-2 were detected by RT-PCR. Results: The time of tumorigenesis in the experimental group was delayed, the growth rate was slowed down, the volume and weight of the transplanted tumor were significantly reduced (P <0.05), the atypia of the tumor was reduced and the necrosis was significantly increased by HE staining. Immunohistochemistry and Western blot showed that the HIF- (P <0.05). The mRNA expression of HIF-1α, EPHA2, VE-cadherin and MMP2 in experimental group were all decreased to different extents, and the expression of EPHA2 and VE- VE-cadherin decreased significantly (P <0.05). Conclusion: HIF-1α silencing can significantly inhibit the growth of transplanted esophageal squamous carcinoma xenografts in nude mice and can effectively inhibit the expression of vasculogenic mimic gene EPHA2 and VE-cadherin in xenograft tumors. HIF-1α may be involved in these two sites Regulation of Angiogenesis Mimicry in Esophageal Squamous Cell.