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目的 :观察血管内皮生长因子16 5 (VEGF16 5 )反义RNA对人食管鳞癌细胞EC10 9的影响 ,探讨其治疗食管癌的可行性。方法 :采用亚克隆技术 ,构建并鉴定VEGF16 5 反义RNA的真核表达载体。以重组质粒转染人食管鳞癌细胞EC10 9后 ,将其接种于裸鼠皮下 ,分别利用原位杂交、激光共聚焦、图象分析及微血管计数等方法 ,观察转染前后EC10 9细胞的生物学性状和致瘤性。结果 :成功地构建了VEGF16 5 反义RNA的真核表达载体 ,并在EC10 9细胞中获得表达。转染细胞中VEGF16 5 的表达下降 75% ,其生物学性状不受外源基因表达的影响 ,但其在裸鼠皮下的致瘤性和和瘤组织中血管的生成明显下降。VEGF16 5 反义RNA转染组、空载体转染组和对照组中肿瘤的体积 ,分别为 (82 0± 112 .5)mm3 、(793 0± 10 3 5)mm3 和 (7850± 950 )mm3(P <0 .0 1) ;微血管的密度分别为 (8.5± 1.2 ) /mm2 、(44.3± 9.4) /mm2 和 (46.4± 12 .6) /mm2 (P <0 .0 1)。结论 :VEGF16 5反义RNA能够明显减少食管鳞癌细胞内VEGF16 5 的表达 ,具有抑制肿瘤生长和血管生成的作用 ,可望用于实体肿瘤的辅助治疗。
OBJECTIVE: To observe the effect of VEGF165 antisense RNA on human esophageal squamous carcinoma cell line EC109 and explore the feasibility of its treatment for esophageal cancer. Methods: The subclone technique was used to construct and identify the eukaryotic expression vector of VEGF16 5 antisense RNA. The recombinant plasmids were transfected into human esophageal squamous carcinoma cell line EC109 and then inoculated into nude mice subcutaneously. The in vivo hybridization, confocal laser scanning microscopy, image analysis and microvessel counting were used to observe the growth of EC109 cells Learn traits and tumorigenicity. Results: The eukaryotic expression vector of VEGF16 5 antisense RNA was successfully constructed and expressed in EC10 9 cells. The expression of VEGF165 in transfected cells decreased by 75%. The biological characteristics of the transfected cells were not affected by the expression of exogenous gene. However, the tumorigenicity of VEGF165 and the formation of blood vessels in tumor tissue were significantly decreased. The volume of tumor in VEGF165 antisense RNA transfection group, empty vector transfection group and control group were (82 ± 112.5) mm3, (793 ± 1035) mm3 and (7850 ± 950) mm3 (P <0.01). Microvessel density was (8.5 ± 1.2) / mm2, (44.3 ± 9.4) / mm2 and (46.4 ± 12.6) / mm2 respectively (P <0.01). CONCLUSION: VEGF165 antisense RNA can significantly reduce the expression of VEGF165 in esophageal squamous cell carcinoma cells, and can inhibit tumor growth and angiogenesis. It is expected to be used in the adjuvant therapy of solid tumors.