Abstract
Cancer is the second leading cause of death in the world. About 18 milllion people were affected with cancer in 2018. Research in novel anti-cancer agents of bacterial origin thus gain importance. Azurin is a novel bacterial protein that has unique ability to specifically target cancer cells without any harmful effect to normal cells. In this study, cytotoxic effect of azurin from Pseudomonas aeruginosa SSj expressed in pET22b (+) vector and standard azurin (Sigma) were studied on (MCF7), (HCT15), and (VERO) cell lines by MTT assay. Apoptosis assay was performed using Hoechst staining and confirmed by FRET based assay on colon cancer cell lines (SW480). In vivo ascites antitumor model assay was also done to detect the in vivo cytotoxicity of azurin. Anti-hemolytic potential of purified azurin on human erythrocytes was also evaluated. Azurin induced cytotoxicity in both HCT15 and MCF7 cell lines, without having any adverse effect on the normal cells. Cell cycle analysis by Hoechst staining and FRET analysis, both showed evidence of apoptosis induced by azurin. In vivo ascites antitumor model proved that azurin treatment can increase the lifespan of tumor-bearing mice. Anti-hemolytic assay showed a significant increase of inhibition of hemolysis.
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Abbreviations
- MCF7:
-
Michigan Cancer Foundation-7
- HCT 15:
-
Human colorectal carcinoma
- SW480:
-
Spectral karyotyping of the human colon cancer cell line
- MTT:
-
3-(4,5-Dimethylthiazol-2-Yl)-2,5-diphenyl tetrazolium bromide
- DMEM:
-
Dulbecco’s modified Eagle medium
- FBS:
-
Fetal bovine serum
- PBS:
-
Phosphate-buffered saline
- DLA:
-
Daltons lymphoma ascites
- EYFP:
-
Enhanced yellow fluorescent protein
- ECFP:
-
Enhanced cyan fluorescent protein
- DAPI:
-
4′,6-Diamidino-2-phenylindole
- FSC:
-
Forward scatter
- SSC:
-
Side scatter
- FRET:
-
Fluorescence resonance energy transfer
- DEVD:
-
Aspartic acid–glutamic acid–valine–aspartic acid
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Sereena, M.C., Sebastian, D. Evaluation of Anticancer and Anti-hemolytic Activity of Azurin, a Novel Bacterial Protein from Pseudomonas aeruginosa SSj. Int J Pept Res Ther 26, 459–466 (2020). https://doi.org/10.1007/s10989-019-09851-1
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DOI: https://doi.org/10.1007/s10989-019-09851-1