毕赤酵母表达重组人内皮抑素对小鼠肺腺癌LA795生长和转移的抑制作用

背景与目的：肿瘤的生长和转移有赖于新生血管的生成,内皮抑素能抑制肿瘤的血管生成。本研究旨在观察毕巴斯德酵母（pichia.pastoris.GS115)分泌表达的重组人内皮抑素（recombinant human endostatin,rhES)对小鼠肺腺癌LA795生长和转换的抑制作用。方法：挑取一株高效分泌表达rhES的毕赤巴斯德酵母菌株,利用甲醇进行大量诱导表达;用肝素亲和层析的方法纯化目的蛋白;将接种LA795肺腺癌细胞的T739小鼠随机分成两组,分别给予rhES和PBS皮下注射,每日1次,连续14天;观察两组小鼠肿瘤生长情况,测量肿瘤体积大小,并观察两组小鼠肿瘤肺部转移情况。结果：经甲醇诱导的毕赤巴斯德酵母菌株高效分泌表达了rhES;动物实验研究发现rhES能显著抑制小鼠肺腺癌LA795的生长,rhES治疗组肿瘤大小与对照组比较具有显著性差异（P＜0．001）,抑瘤率达到66．4％,并且有效抑制了肿瘤的肺部转移。结论：利用毕赤酵母作为宿主分泌表达的rhES具有良好的生物学活性,可显著抑制小鼠肺腺癌LA795的生长和转移。

Inhibitory effects of recombinant human endostatin on growth and metastasis of lung adenocarcinoma LA795 in mice

Background & Objective:Generally, growth and metastasis of tumor are critically dependent on angiogenesis. Endostatin can specifically inhibits tumor angiogenesis. The current study was designed to evaluate the inhibitory effect of recombinant human endostatin (rhES) secreted by pichia.pastoris.GS115 on the growth and metastasis of mice lung adenocarcinoma LA795 in mice T739. Methods:To select a strain that could highly express recombinant human endostatin, then induce the clone to express rhES by adding methanol. Purification of rhES was performed with heparin affinity chromatography. LA795 tumor cells were inoculated subcutaneously into the dorsa of T739 mice, and the mice were randomized into two groups. The first group was given rhES(20 mg/kg/d), and the second group was given equal volume of PBS, for 14 consecutive days. The volume of tumors were measured. And the tumor metastasis in the lungs of the mice was observed. Results: The selected clone was induced to secrete enough soluble rhES. The purified protein could strongly inhibit growth and metastasis of mice lung adenocarcinoma LA795 in T739 mice (P< 0.001). Conclusion: The rhES secreted by pichia. pastoris.GS115 has good biological activities and greatly inhibit growth and metastasis of mice lung adenocarcinoma LA795 in mice T739.