Granzyme B-truncated VEGF fusion protein represses angiogenesis and tumor growth of OSCC
Granzyme B-truncated VEGF fusion protein represses angiogenesis and tumor growth of OSCC
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颗粒酶 B 截短的 VEGF 融合蛋白抑制 OSCC 的血管生成和肿瘤生长
DOI：
10.1111/odi.12522
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
X-Z lv
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作者：
X-Z lv
目的研究人颗粒酶B（human granzyme B，hGrB）与截短型血管内皮生长因子（truncated vascular endothelial growth factor，tVEGF）融合蛋白hGrB-TV对人口腔鳞状细胞癌（oral squamous cell carcinoma，OSCC）的体外和体内抗肿瘤作用。体外检测hGrB-TV对VEGFR-2（Flk-1）+OSCC细胞的细胞毒作用。结果纯化的hGrB-TV融合蛋白可选择性地内化入VEGFR-2（Flk-1）+OSCC细胞和内皮细胞中。它可以将失活的caspase 3切割成其活性p20形式。hGrB-TV对VEGFR-2+SCC-9细胞显示出剂量依赖性细胞毒性。细胞在十几分钟内出现形态学改变和细胞溶解。然而，在VEGFR-2−细胞上未观察到细胞毒性。单独的hGrB或单独的tVEGF对SCC-9细胞没有任何毒性。此外，hGrB-TV处理完全破坏了体内鸡胚绒毛尿囊膜（CAM）的血管系统，从而导致鸡胚发育停滞。最重要的是，融合蛋白hGrB-TV抑制肿瘤血管生成和生长的人口腔鳞癌裸鼠移植瘤没有任何明显的toxicsConclusionsThe融合蛋白hGrB-TV特异性抑制血管生成和肿瘤生长的口腔鳞癌; hGrB-TV是一个强大的和安全的治疗分子，用于肿瘤治疗。
ObjectiveTo evaluate the antitumor effects of fusion protein hGrB‐TV of human granzyme B (hGrB) and truncated vascular endothelial growth factor (tVEGF) on human oral squamous cell carcinoma (OSCC)in vitroandin vivo.MethodsThe fusion protein hGrB‐TV was expressed and purified fromE. colibacteria by affinity chromatography. The cytotoxcity of hGrB‐TV on VEGFR‐2 (Flk‐1)+OSCC cells was analyzedin vitro. The antitumor therapeutic study was conducted on OSCC xenograftsin vivo.ResultsThe purified hGrB‐TV fusion protein was selectively internalized into VEGFR‐2 (Flk‐1)+OSCC cells and endothelial cells. It can cleave inactive caspase 3 into its active p20 form. The hGrB‐TV showed dose‐dependent cytotoxicity on VEGFR‐2+SCC‐9 cells. The morphological changes and cytolysis were appeared within dozen minutes. However, no cytotoxicity was observed on VEGFR‐2−cells. The hGrB alone or tVEGF alone did not have any toxicity on SCC‐9 cells. In addition, hGrB‐TV treatment completely destroyed the vasculature of the chick chorioallantoic membrane (CAM)in vivoand consequently led to chick embryo development arrest. Most importantly, the fusion protein hGrB‐TV inhibited tumor angiogenesis and growth of human OSCC xenografts in nude mice without any apparent toxicity.ConclusionsThe fusion protein hGrB‐TV specifically inhibits angiogenesis and tumor growth of OSCC; hGrB‐TV is a powerful and safe therapeutic molecule for tumor therapy.