目的：研究血管内皮生长因子(VEGF)反义RNA对人食管癌细胞的抑制作用，探讨以VEGF反义RNA进行食管癌基因治疗的可行性。方法：采用脂质体法将反义VEGFc DNA质粒转染人食管癌细胞TE-1；用MTT法检测细胞增殖情况；用免疫组化法检测VEGF蛋白表达；原位杂交和RT—PCR技术检测VEGFmRNA的表达。流式细胞术检测细胞的凋亡率和周期分布。结果：被反义VEGFcDNA质粒转染的食管癌细胞有外源性VEGF反义基因的整合及表达，该细胞的VEGFmRNA及蛋白表达水平降低，但细胞生长速率无明显改变，未发生明显凋亡现象；接种裸鼠后，形成肿瘤的时间明显延长，肿瘤的体积和重量明显减小。结论：VEGF反义RNA可抑制食管癌细胞VEGF表达和裸鼠体内的肿瘤生长。该表达体系为食管癌基因治疗的进一步研究奠定了基础。

Objective: To investigate the inhibitory effect of VEGF antisense RNA on the growth of human esophageal cancer cells in vitro, in order to further investigate the feasibility of VEGF antisense RNA gene therapy of esophageal cancer. Methods: The plasmid carrying with VEGF antisense cDNA was transfected into esophageal cancer cells, and confirmed its expression by RT-PCR. The expression level of VEGFmRNA and VEGF protein was examined in antisense group by insitu hybridization and immunohistochemistry staining. The cell growth rate was detected by MTT assay. Apoptotic rate in transfected cells was detected by FCM assay. Results: The expression of exogenous antisense VEGFmRNA was confirmed in transfected cells, and the VEGF protein and endogenous VEGFmRNA were dramatically decreased. The growth rate of transfected cells was not inhibited. Apoptotic cells were not found in transfected cells. Latent period of the tumor formation of the antisense group was lengthened, while body weight, volume of tumors was significantly smaller than that of empty vector group and control group. Conclusions: VEGF antisense RNA could decrease the expression of VEGF of e-sophageal cancer and cell proliferation in vivo, which may apply a useful theory basis for gene therapy of esophageal canc-er.

R730.54

山东省重大攻关项目（NO. 31050102）