重组血管抑制剂CHM-I对骨肉瘤的治疗作用

Therapeutic effect of recombinant angiogenic inhibitor CHM- I on osteosarcoma

目的:观察重组人软骨调节素(chondromodulin,CHM)蛋白对骨肿瘤的治疗作用。方法:利用RT-PCR技术从人软骨组织中扩增出CHM-I基因片段,并将其克隆到原核表达载体pET28a,在大肠杆菌中诱导表达和纯化重组人软骨调节素蛋白,将重组蛋白与人脐静脉上皮细胞或MG-63骨肉瘤细胞共培养,观察其对内皮细胞形成管样结构和MG-63骨肉瘤细胞生长的影响;将重组蛋白局部注射裸鼠皮下肿瘤,观察其对肿瘤形成的影响。结果:获得纯化的重组人软骨调节素蛋白,该蛋白在体外可以抑制血管内皮细胞管样结构的形成而对肿瘤细胞的生长无影响;重组蛋白瘤内注射可以抑制肿瘤在裸鼠体内的生长。结论:重组人软骨调节素蛋白通过抑制血管形成而抑制肿瘤生长,具有良好的临床应用价值。

Objective： To study the effects of recombinant human Chondromodulin-I protein in treatment of osteosarcoma. Methods： Human Chondromodulin-I gene was amplified from cartilage tissue by RT-PCR and was cloned into procaryotic expression vector Pet28a（ ＋ ）. The recombinant plasmid Pet（CHM-I） was transformed into E. coli B121 （DE3） ; the product was fused with 6 x His at N-terminal and was purified by Ni2 ＋ 2NTA ion exchange resin. The formation of tube-like cellular networks was assayed by co-incubation of HUVECs and recombinant human Chondromodulin-I. The proliferation of the cells incubated with the recombinant human Chondromodulin-I protein was examined by MTT assay and cell number counter Null mice received local injection of recombinant human Chondromodulin-I protein into the tumors. Results： Rcombinant human CHM-I protein was successfully obtained. The purified CHM-I did not inhibit the proliferation of tumour cells, but inhibited the formation of tubelike cellar network in vitro and tumor growth. Conclusion： Recombitant human Chondromodulin-I protein can inhibit angiogenesis in vitro and growth of tumor in vivo