metallopeptidase gene under colon-specific promoter on the colorectal and breast cancer cell lines.

2021-04-01
Iranian journal of basic medical sciences 24(4)
- Narges Dadfarma
- Jamileh Nowroozi
- Bahram Kazemi
- Mojgan Bandehpour

PubMed: 34094033
DOI: 10.22038/ijbms.2021.53015.11950

Objectives

Anti-tumor effects of Lactobacilli as normal flora have been described. In a previous study, we identified a protein isolated from the bacterium Lactobacillus casei ATCC 39392 in acidic pH conditions named metallopeptidase. Therefore, we decided to evaluate the effect of the recombinant plasmid coding metallopeptidase protein on the inhibition, proliferation, or apoptosis of the colorectal and breast cancer cell lines.

Materials and methods

Identified metallopeptidase gene of L. casei under the specific colon cancer promoter was transferred to the Human SW480 and MDA-MB231 cells. Cell viability was evaluated in these two cancer cell lines via MTT assay, apoptotic changes, and expression level of p53 and MAP2K1 genes in comparison with healthy blood cells as a control group.

Results

Viability of SW480 and MDA-MB231 cells was identified at 25% and 7%, respectively. An increase in apoptotic cell death in the SW480 cell line was observed as revealed by Tunnel staining. The expression assay of TP53 and MAP2K1 genes showed that MPL protein altered gene expression in a cell type-specific manner. Tunnel analyses showed that the pronounced cytotoxic effect of pEGFP-C2/MPL plasmid on SW480 cells was mediated through apoptosis.

Conclusion

These results suggest that endogenous recombinant MPL under colon specific promoter inhibits the proliferation of SW480 colorectal cancer cells by increase in MAP2K1 and P53 activation. L. casei metallopeptidase under the same circumstances could not affect the growth rate and viability of MDA-MB231 breast cancer cells in vitro.

Research Insights

Supplement	Health Outcome	Effect Type	Effect Size
Lactobacillus brevis SBC8803	Increased Apoptosis	Beneficial	Moderate
Lactobacillus brevis SBC8803	Increased Apoptosis in Cancer Cells	Beneficial	Moderate
Lactobacillus brevis SBC8803	Reduced Cancer Cell Viability	Beneficial	Large
Lactobacillus brevis SBC8803	Reduced Colon Cancer Cell Proliferation	Beneficial	Large
Lactobacillus brevis SBC8803	Reduced Colon Cancer Cell Viability	Beneficial	Large