【作者】 杨进强； 李勇； 赵增仁； 王益民； 邝钢； 范立侨； 赵群；

【Author】 Yang Jin-Qiang Li Yong Zhao Zeng-Ren et al The Third Department of Surgery, the Fourth Hospital of Hebei Medical University & Hebei provincial tumor hospital, Shijiazhuang 050011, China

【机构】 河北医科大学第四医院暨河北省肿瘤医院外科；

【摘要】 目的:探讨脂质体转染VEGF-C反义寡核苷酸在人胃癌OCUM-2MD3细胞中的转染效率及对VEGF-C表达的影响。方法:以VEGF—C基因翻译起始区为靶点合成反义寡核苷酸,脂质体介导转染人胃癌OCUM-2MD3细胞,荧光显微镜观察转染效率,RT—PCR检测转染后VEGF— CmRNA表达水平,流式细胞术检测VEGF-C蛋白表达。结果:经脂质体介导可成功将VEGF—C反义寡核苷酸转染至胃癌OCUM-2MD3细胞,12小时达其峰值87．93％。反义组 VEGF—CmRNA相对吸光度值为0．38±0．11,与正义组(0． 83±0．17)和对照组(0．80±0．14)比较,表达水平显著下调(P <0．01)。反义组VEGF—C蛋白表达水平(31．26±5．17,)％, 较正义组(41．83±7．49)％和对照组(38．65±7．11)％亦明显下降,差异有显著性(P<0．01)。结论:VEGF-C反义寡核苷酸经脂质体介导转染胃癌OCUM-2MD3细胞可降低胃癌细胞VEGF-CmRNA及其蛋白表达水平。更多还原

【Abstract】 Obleotlve: To investigate the influence of VEGF-C antisense oligodeoxynuclecotide (ASODN) on the VEGF-C expression and transfection effects in human gastric cancer cell OCUM-2MD3 induced by liposome. Methods: The ASODN sequence, which was synthesized according to the coding region of VEGF-C gene, was transformed into human gastric cancer cell OCUM-2MD3 induced by liposome. The transfection effects were observed by fluorescence microscope. The changes in the expression of VEGF-C mRNA and protein were determined by RT-PCR and flow cytometry(FCM), respectively. Results: VEGF-C-ASODN could be successfully transformed into gastric cancer cell induced by liposome, and its optimal ratio was 87.93% after 12 hours. Compared with the control and oligodeoxynuclecotide (SODN) group, not only was the VEGF-C mRNA level (Ar=0.38 ± 0.11) significantly decresaed(P < 0.01), but also the protein of VEGF-C (31.26 ± 5.17)% was dramatically lowed(P < 0.01). Conclusion: VEGF-C-ASODN that was transformed into human gastric cancer cell induced by liposome may inhibit the expression of VEGF-C mRNA and protein.更多还原