关键词:

ETAR 羧基末端, 原核表达, 黑色素瘤, 细胞迁移, 细胞侵袭

Abstract:

Objective To obtain the 58 amino acid peptide of the carboxyl terminal of ETAR（ETAR ⁃ C58）and investigate the effects on the migration and invasion of melanoma A375 cells. Methods ETAR ⁃C58 sequence was amplified by PCR，inserted into the vector pWaldo⁃GFPe and then transformed into Escherichia coli BL21（DE3）. After expression，the fusion peptide was purified by nickel column affinity chromatography and gel filtration chromatography，analyzed by SDS ⁃PAGE and further identified by immunoblotting. Finally，Transwell assay was carried out to measure the migration and invasion of A375cells. Results The recombinant expression vector pWaldo⁃ETAR⁃C58 was successfully constructed and the soluble fusion peptide was accounted for approxi⁃ mately 90%. SDS⁃PAGE and immunoblotting results verified the successful purification of the ETAR⁃C58 with nearly 95% purity. ETAR ⁃ C58 peptide could inhibit A375 cell migration and invasion at 2 μmol/L. Conclusion The ETAR⁃C58 peptide have anti⁃melanoma activity，which allows a possibility for further clinical application.

Key words:

ETAR carboxyl terminal, prokaryotic expression, melanoma, cell migration, cell invasion