Proliferative Effect of FadA Recombinant Protein from Fusobacterium nucleatum on SW480 Colorectal Cancer Cell Line

Colorectal Cancer
21/07/2020

Infect Disord Drug Targets. 2020 Jul 19. doi: 10.2174/1871526520666200720113004. Online ahead of print.

ABSTRACT

BACKGROUND AND AIM: Colorectal Cancer (CRC) is one of the most frequent cancers diagnosed in both men and women worldwide. Fusobacterium nucleatum adhesin A (FadA) has an important potential factor in the development or progression of CRC. The aim of the present study was to evaluate the proliferative effect of recombinant FadA on SW480 colorectal cancer cell line.

MATERIALS AND METHODS: The recombinant pET21(b)-fadA plasmid was synthesized and transformed into competent E.coli DH5α. In the next step, induction and expression of recombinant FadA were carried out in E. coli BL21 (DE3) competent cells. Expression and purification of protein were successfully done and it was analyzed and confirmed by SDS-PAGE and western blotting. The proliferative effect of purified FadA on SW480 CRC cell line was evaluated using MTT assay and cell counting methods.

RESULTS: Visualization of the specific band isolated from the linear plasmid on the agarose gel confirmed the presence of the desired gene. After electrophoresis and Coomassie blue staining, the protein of interest with an approximate molecular weight of 13KDa was detected. The MTT assay, similar to cell counting methods, revealed that FadA dose and timedependently promoted SW480 cell growth and proliferation in 24, 48 and 72 hours.

CONCLUSION: The results showed that FadA stimulates proliferation of SW480 colorectal cancer cell line with a dose and time-dependent manner.