基因重组心肌肌钙蛋白I融合蛋白抗肿瘤作用实验研究

The Experimental Study of Tumor Growth of Recombinant Protein Fused with Cardiac Troponin I and Artificial Peptide

摘要: 目的：研究基因重组心肌肌钙蛋白I 与人工短肽融合蛋白(CIS)对肿瘤生长的抑制作用。方法：用Western Blot法观察对人肝癌细胞MHCC97H的VEGF-A和VEGF-C、MMP-9的影响。用3种人肿瘤异位可移植瘤模型观察CIS在裸鼠体内对肿瘤生长的抑制作用。结果：CIS对MHCC97H细胞，CIS 200 mg/ml，48 h，对VEGF-A表达较肿瘤模型组显著减少，抑制率为23.8%。DDP 2 mg/ml，48 h，对VEGF-A表抑制率为25.8%。CIS 200 mg/ml，48 h，对MMP-9表达较肿瘤模型组减少显著，抑制率为18.1%，DDP 2 mg/ml，48 h，对MMP-9表达比较肿瘤模型组抑制率为12.6%。CIS H组和DDP组对VEGF-C表达抑制作用较弱。荷瘤鼠体内皮下移植瘤模型实验显示CIS (10 mg/kg，SC)处理组肿瘤生长缓慢，瘤体明显小于模型对照组，对人肝癌HCC9204肿瘤瘤重抑制率为54.13%，对人可移植性乳腺癌BCaP-37肿瘤瘤重抑制率为53.88%，对人可移植性结肠癌HCT116肿瘤瘤重抑制率为41.12%。结论：基因重组心肌肌钙I与人工短肽融合蛋白CIS对人肝癌细胞MHCC97H VEGF-A和MMP-9表达具有抑制作用。在体内，CIS融合蛋白可有效的抑制人可移植肿瘤细胞的生长，CIS抗肿瘤效应与抑制肿瘤组织中血管内皮生长因子的表达和人基质金属蛋白酶-9的表达有关。

Abstract: Objective: The effect of antitumor growth of recombinant cardiac troponin fusion protein com-posed of subunit I and artificial peptide, called CIS, was examined in this study. Methods: The CIS’s effect on the expressing of human vascular endothelial growth factor A (VEGF-A) was examined using Western blot assay in vitro, and matrix metallopeptidase 9 was studied with purified recombinant CIS protein. The effect of tumors growth treated with CIS was performed using several in vivo nude mice xenograft models. Results: There was an evident reduction on VEGF-A, MMP-9 when the human hepatocellular carcinoma cells MHCC97H were treated with purified CIS fusion protein, which was also shown in dose-dependent manner compared to the untreated control group. A significant inhibition of tumor growth rate was achieved in CIS treated nude mice compared to untreated control mice in 8 or 6 different nude mice xenograft models. Conclusions: The fusion protein CIS has shown the inhibitive effect on the tumor growth in our in vivo nude mice models, which could be mediated by the mechanism of CIS’s effect on the inhibition of VEGF-A, MMP-9. This work has built the foundation for the in-depth investigations on the CIS’s pharmaceutical application targeting the anti-tumor therapy.