CAJ | 학술논문

背景：胶质瘤的根除治疗至今仍是一大难题，抗血管生成治疗胶质瘤有望成为新的有效途径。 　　目的：证实内皮抑素在体外对血管生长的抑制作用，为今后利用其抑制肿瘤生长奠定实验室基础。 　　方法：取Wistar大鼠肝脏，提取mRNA后利用RT-PCR获取内皮抑素cDNA片段。碱裂解法小量提取质粒pcDNA3。重组质粒 pcDNA3-Endo 的构建。重组 pcDNA3-Endo 真核表达载体转染骨髓间充质干细胞。RT-PCR及Western Blot检测内皮抑素基因的表达。MTT法检测ECV-304细胞增殖抑制实验。体外实验共分成4个组：重组质粒组、空载质粒组、脂质体对照组及空白对照组。 　　结果与结论：成功构建pcDNA3-Endo重组真核表达质粒，pcDNA3-Endo质粒能在体外有效转录并分泌内皮抑素基因，转染了外源pcDNA3-Endo质粒的ECV-304细胞增殖明显受到抑制。结果提示内皮抑素基因能在体外有效抑制血管内皮细胞的增殖。
배경：효질류적근제치료지금잉시일대난제，항혈관생성치료효질류유망성위신적유효도경。 　　목적：증실내피억소재체외대혈관생장적억제작용，위금후이용기억제종류생장전정실험실기출。 　　방법：취Wistar대서간장，제취mRNA후이용RT-PCR획취내피억소cDNA편단。감렬해법소량제취질립pcDNA3。중조질립 pcDNA3-Endo 적구건。중조 pcDNA3-Endo 진핵표체재체전염골수간충질간세포。RT-PCR급Western Blot검측내피억소기인적표체。MTT법검측ECV-304세포증식억제실험。체외실험공분성4개조：중조질립조、공재질립조、지질체대조조급공백대조조。 　　결과여결론：성공구건pcDNA3-Endo중조진핵표체질립，pcDNA3-Endo질립능재체외유효전록병분비내피억소기인，전염료외원pcDNA3-Endo질립적ECV-304세포증식명현수도억제。결과제시내피억소기인능재체외유효억제혈관내피세포적증식。
BACKGROUND:The eradication therapy of glioma is the major problem, and anti-angiogenesis therapy is a potential treatment of glioma. OBJECTIVE:To confirm the inhibiting effect of endostatin on angiogenesis in vitro, and to lay the foundation in inhibiting the growth of tumor by endostatin in the future. METHODS:Endostatin mRNA was extracted from the liver of Wistar rats by Trizol and endostatin cDNA was synthesized by RT-PCR. Endostatin cDNA and pcDNA3 were connected and pcDNA3-Endo recombined plasmid was constructed successful y. The recombinant pcDNA3-Endo was transfected into bone marrow mesenchymal stem cells by Lipofectamine. The expression of endostatin was identified by RT-PCR and western blot analysis. Endostatin proteinum activity was detected by ECV-304 cellproliferation inhibition experiment using MTT assay. The in vitro experiments were divided into four groups:recombinant plasmid group, vector plasmid group, liposome control group and blank control group. RESULTS AND CONCLUSION:PcDNA3-Endo eukaryon expression plasmid was constructed successful y. Endostatin gene can be transcribed and expressed effectively in vitro by pcDNA3-Endo plasmid. The growth of ECV-304 cellwas inhibited obviously by pcDNA3-Endo. The growth of vascular endothelial cells can be inhibited obviously by endostatin gene.