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Tumor-targeted induction of intrinsic apoptosis in colon cancer cells by Lactobacillus plantarum and Lactobacillus rhamnosus strains

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Abstract

Background

Colorectal cancer is one of the widespread and lethal types of malignancies. Recently, antineoplastic attributes of probiotics have attracted lots of attention. Here, we investigated anti-proliferative potential of the non-pathogenic strains Lactobacillus plantarum ATCC 14,917 and Lactobacillus rhamnosus ATCC 7469 on human colorectal adenocarcinoma-originated Caco-2 cells.

Methods and results

Caco-2 and HUVEC control cells were treated with ethyl acetate extracts of the two Lactobacillus strains to assess cell viability by MTT assay. Annexin/PI staining flow cytometry, and caspase-3, -8 and − 9 activity assays were performed to determine the type of cell death induced in extract-treated cells. Expression levels of apoptosis-related genes were evaluated by RT-PCR. Extracts from both L. plantarum and L. rhamnosus specifically targeted the Caco-2 cells and not HUVEC controls, and significantly affected the viability of the colon cancer cell line in a time- and dose-dependent manner. This effect was shown to occur through activation of the intrinsic apoptosis pathway, as indicated by the increased caspase-3 and − 9 activities. While there are limited and conflicting data about the mechanisms underlying the specific antineoplastic attributes of Lactobacillus strains, we clarified the overall induced mechanism. The Lactobacillus extracts specifically down-regulated the expression of the anti-apoptotic bcl-2 and bcl-xl, and simultaneously up-regulated the pro-apoptotic bak, bad, and bax genes in treated Caco-2 cells.

Conclusions

Ethyl acetate extracts of L. plantarum and L. rhamnosus strains could be considered as targeted anti-cancer treatments specifically inducing the intrinsic apoptosis pathway in colorectal tumor cells.

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Abbreviations

Bcl-2::

B-cell lymphoma 2

Bcl-xL::

B-cell lymphoma-extra, long isoform

BAD::

Bcl-2 associated agonist of cell death

BAK::

Bcl-2 homologous antagonist killer

BAX::

Bcl-2-associated X protein

DMEM::

Dulbecco’s Modified Eagle medium

EPS::

Crude exopolysaccharide

IECs::

Intestinal epithelial cells

L. plantarum::

Lactobacillus plantarum

L. rhamnosus::

Lactobacillus rhamnosus

LGG::

L. rhamnosus GG CGMCC 1.2134

MRS::

De Man, Rogosa and Sharp

MTT::

3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide

PI::

Propidium Iodide

pNA::

p-nitroaniline

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Funding

The authors did not receive support from any organization for the submitted work. No funding was received to assist with the preparation of this manuscript. No funding was received for conducting this study. No funds, grants, or other support was received.

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Authors and Affiliations

  1. Department of Microbiology, Cellular and Molecular Research Center, Infectious and Tropical Diseases Research Center, Health Research Institute, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

    Mansour Amin, Tahereh Navidifar & Sholeh Saeb

  2. Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

    Ebrahim Barzegari

  3. Department of Biochemistry, Abadan University of Medical Sciences, Abadan, Iran

    Mostafa Jamalan

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  1. Mansour Amin
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  2. Tahereh Navidifar
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Contributions

In this research, Mansour Amin, Tahereh Navidifar, Sholeh Saeb, Ebrahim Barzegari, and Mostafa Jamalan have been in charge of research, data collection and analysis, and article writing.

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Correspondence to Mostafa Jamalan.

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This research project was approved by the Ethics Committee of Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

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Amin, M., Navidifar, T., Saeb, S. et al. Tumor-targeted induction of intrinsic apoptosis in colon cancer cells by Lactobacillus plantarum and Lactobacillus rhamnosus strains. Mol Biol Rep 50, 5345–5354 (2023). https://doi.org/10.1007/s11033-023-08445-x

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