摘要
目的研究经RGD修饰的人内皮抑素(endostatin)N-端的30肽对胃癌SGC-7901细胞增殖及转移能力的影响。方法人工合成人内皮抑素1～30位氨基酸(30肽,25～31序列由RGIRGAD改为RGDRGD)所对应的核苷酸序列,连接到质粒pTYB2中,再转化至大肠杆菌BL21(DE3)中表达,同法合成人内皮抑素1～27位氨基酸构成的27肽作为对照。用WST-1试剂检测SGC-7901细胞的增殖情况;Transwell实验检测细胞迁移和侵袭能力;明胶酶谱实验观察30肽和27肽对基质金属蛋白酶MMP2和MMP9活性的影响。结果与27肽相比,30肽能够有效抑制SGC-7901细胞的增殖、侵袭和迁移,同时抑制了MMP2和MMP9的活性。结论 30肽可能是通过下调MMP2和MMP9的活性而抑制SGC-7901细胞的转移,其在临床胃癌的治疗上具有潜在的应用前景。
Objective The effect of a 30 - amino - acid N - terminal peptide of endostatin （ aa 1 - 30, peptide 30） modified by RGD on proliferation and metatasis of SGC - 7901 cells was studied. Methods The nucleotidc sequence encoding amino acids 1 - 30 of endostatin （peptide 30, with amino acids 25 -31 mutated from to RGDRGD） was artificially synthesized and cloned into the plasmid pTYB2 and expressed in E. coli （DE3）. Similarly, peptide 27, corresponding to amino acids 1 - 27 of endostatin, was produced as a control. The proliferation of SGC - 7901 was observed by WST - 1 kit. Cell migration and invasion ablity were examined by Transwell. The activity of MMP - 2 （ metalloproteinase - 2） and MMP - 9 （ metalloproteinase - 9） were determined by gelatin zymography. Results Pep- tide 30 effectively suppressed the proliferation and invasion and migration abilities of SGC -7901cells. Moreover, peptide 30 inactivated MMP - 2 and MMP - 9 activity. Conclusion The mechanism of peptide 30 suppressed cancer metastasis may involve the down - regula- tion of MMP - 2 and MMP - 9 activity. The peptide 30 can potentially be used to treat gastric carcinoma in clinic.
出处
《医学研究杂志》
2013年第2期96-100,共5页
Journal of Medical Research
基金
黑龙江省教育厅基金资助项目(12511619)
