Recombinant Expression of hTFF2 and Its Roles in Migration and Anchorage-independent Growth of Colonic Cancer Cells
- VernacularTitle:重组表达hTFF2促进结肠癌细胞迁移和失巢增殖作用
- Author:
Weiguo JIANG
;
Fangmei LUO
;
Haoying WANG
;
Wei XIE
;
Yu QIN
;
Youguang HUANG
- Publication Type:Journal Article
- Keywords:
Trefoil factor 2;
Colonic cancer;
Cell migration;
Anchorage-independent growth
- From:
Journal of Kunming Medical University
2013;(8):31-35
- CountryChina
- Language:Chinese
-
Abstract:
Objective To over-express human trefoil factor 2 (hTFF2) by Escherichia coli system and an-alyze its activities in promoting migration and anchorage-independent growth in SW480 colonic cancer cells. Meth-ods hTFF2 gene encoding mature peptide was obtained by RT-PCR, and the recombinant expression vector pET32a-hTFF2 was constructed. Then pET32a-hTFF2 was transformed into E. coli BL21-32a and TrxA-hTFF2 fu-sion protein was induced to over-express. The expressed product was isolated by Ni-NTA affinity chromatography, purified by dialysis and identified by Western blotting. The activities of the recombinant hTFF2 in promoting SW480 cells migration and anchorage-independent growth were analyzed by MicroChemotaxis Chamber migration assay and Soft-agar assay,respectively. Results The TrxA-hTFF2 fusion protein was expressed to 220 mg/L at high purity. In vitro model demonstrated that recombinant hTFF2 obviously enhanced SW480 cell migration activity and anchor-age-independent growth. Conclusion The recombinant hTFF2 can be expressed in E. coli with high production, purity and biological activities. And its roles in cell migration and anchorage-independent growth suggest that up-regulation of TFF2 in colonic cancer might be involved in cancer invasion and metastases.
